Triazole benzamide derivatives and the compositions and methods of treatment regarding the same

ABSTRACT

The present disclosure is directed to triazole benzamide compounds of formula (I) and formula (II), pharmaceutical compositions thereof and methods for modulating or activating a Parkin ligase The present disclosure is also directed to methods of treating and/or reducing the incidence of diseases or conditions related to the activation of Parkin ligase. R 1 , R 2 , R 3 , M 1 , M 2 , M 3 , L 1 , L 2 , and L 3  are as defined herein.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the priority benefit of U.S. ProvisionalApplication No. 62/345,478, filed Jun. 3, 2016, the disclosure of whichis incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

The present invention relates to triazole benzamide compounds and theirderivatives as well as methods of modulating Parkin ligase or methods oftreating various diseases and conditions with the triazole benzamidecompounds and their derivatives.

BACKGROUND OF THE INVENTION

Ubiquitin-Proteasome Pathway System (UPS) is a critical pathway thatregulates key regulator proteins and degrades misfolded or abnormalproteins. UPS is central to multiple cellular processes, and ifdefective or imbalanced, it leads to pathogenesis of a variety ofdiseases. Posttranslational modification of proteins by ubiquitin is afundamental cellular mechanism that regulates protein stability andactivity and underlies a multitude of functions, from almost everyaspect of biology. The covalent attachment of ubiquitin to specificprotein substrates is achieved through the action of E3 ubiquitinligases. These ligases comprise over 500 different proteins and arecategorized into multiple classes defined by the structural element oftheir E3 functional activity. Specifically, both HECT and RING ligasestransfer an activated ubiquitin from a thioester to the e-amino acidgroup of a lysine residue on a substrate; however, HECT ligases have anactive site cysteine that forms an intermediate thioester bond withubiquitin, while RING ligases function as a scaffold to allow directubiquitin transfer from the E2 to substrate. Recent evidence suggeststhat a subfamily of RING ligases, the RING-between-RING (RBR) family,may contain a catalytic cysteine residue 1,2 in addition to a canonicalRING domain. (Riley et al. 2013. Nat Commun. 4:1982, “Riley et al.”),which is herein incorporated by reference in its entirety.

Deubiquitinating proteins and ubiquitin-specific proteases (DUBs andUSPs) and E3 Ligases play a vital role in the UPS. These proteins aresupported by flexible Zinc Finger (ZnF) domains which stabilize thebinding of ubiquitin (Ub) for specialized functions.

Parkin is a RING-between-RING E3 ligase that functions in the covalentattachment of ubiquitin to specific substrates, and mutations in Parkinare linked to Parkinson's disease, cancer and mycobacterial infection.The individual RING domains for Parkin have been the subject of muchdebate, in regards to the specific residues that coordinate Zn ions, aswell as their relationship to canonical RING crossbrace structuresdefining classical E2-binding domains. R0 is a novel domain structure,but is more similar to Zn-finger domains than to E3 RING domains (Rileyet al. 2013. Nat Commun. 4:1982)

While many drug discovery programs focus on the UPS, few have beensuccessful due to the lack of selectivity and direct access to enzymaticprotein active sites. The present invention is directed towards a novelapproach of disrupting Zn-finger domains that provide a therapeuticbenefit for various diseases and disorders, including oncology andneurology disorders.

SUMMARY OF THE INVENTION

The compounds of the present disclosure can modulate or active Parkinligase and may be useful in treating various diseases and conditions asdisclosed herein. In one embodiment, the present disclosure providescompounds comprising the structure of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L¹, L² and L³ are each independently selected from a bond, alkylene, oralkenylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, —NR⁴C(O)NR⁴—, —C(O)—, —C(═NR⁴)—, —C(═NOR⁴)—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, —NR⁴C(O)O—, —S(O)_(m)—, —S(O)_(m)NR⁴—, or—NR⁴S(O)_(m)—, provided that M¹ and M² are not both —NR⁴—;

R¹, R², and R³ are each independently selected from an alkyl, alkenyl,cycloalkyl, aryl, biphenyl, heterocycloalkyl, heterocyclyl, heteroaryl,cycloalkylalkyl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclylalkyl,heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl, wherein eachcycloalkyl, aryl, heteroaryl, and heterocyclyl portion is optionallysubstituted with one or more R⁵;

R⁴ is each independently H, alkyl, wherein each alkyl is optionallysubstituted with one or more R⁵;

R⁵ is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH,NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶,SO₃H, SO₃R⁶, or SR⁶;

R⁶ is each independently alkyl;

m is 0, 1, or 2; and

wherein the compound is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, L¹, L² and L³ of formula (I) are each independentlyselected from a bond, C1-C3 alkylene, or C2-C3 alkenylene.

In one embodiment, M¹ and M² of formula (I) are each independentlyselected from —NR⁴—, —NR⁴C(O)—, —C(O)NR⁴—, NR⁴C(O)NR⁴—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, or —NR⁴C(O)O—. In another embodiment, M¹and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—.

In one embodiment, R¹, R², and R³ of formula (I) are each independentlyselected from alkyl, aryl, heterocyclyl, heteroaryl, arylalkyl,arylalkenyl, heteroarylalkyl, heteroarylalkenyl, or heterocycloalkyl,wherein each cycloalkyl, aryl, heteroaryl and heterocyclyl portion isoptionally substituted with one or more R⁵. In one embodiment, R⁴ offormula (I) at each occurrence is independently H or C1-C3 alkyl. Inanother embodiment, R¹, R², and R³ are each independently selected fromC1-C3 alkyl, phenyl, 5-10 membered heteroaryl, phenyl-(C1-C3 alkyl),phenyl-(C2-C3 alkenyl), 5-6 membered heteroaryl-(C1-C3 alkyl), orheteroaryl-(C2-C3 alkenyl), wherein each cycloalkyl, aryl, heteroarylportion is optionally substituted with one or more R⁵.

In one embodiment, at least one of R¹, R², and R³ of formula (I) isphenyl-(C2-C3 alkenyl). In another embodiment, at least one of R¹, R²,and R³ is a bicyclic 5-10 membered heteroaryl. In another embodiment, atleast one of R¹, R², and R³ is

In another embodiment, at least one of R¹, R², and R³ is phenyl andsubstituted with at least one of methyl, I, Br, Cl or F.

In one embodiment, a compound of formula (I) has L³ as a bond and R³ asan aryl. In another embodiment, L³ is a bond and R³ is a phenyl.

In one embodiment, -L¹-M¹-R¹ or -L²-M²-R² of a compound of formula (I)are not —CH₂CH₂Ph.

In one embodiment, compounds of formula (I) has the structure of formula(I′):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³, M¹, M², R¹, R², and R³ are as defined for formula (I); and

wherein the compound is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, M¹ and M² of formula (I′) are each independentlyselected from —NR⁴—, —NR⁴C(O)—, —C(O)NR⁴—, NR⁴C(O)NR⁴—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, or —NR⁴C(O)O—.

In one embodiment, R¹, R², and R³ are each independently selected fromalkyl, aryl, heteroaryl, arylalkyl, arylalkenyl, heteroarylalkyl, orheteroarylalkenyl, wherein each cycloalkyl, aryl, heteroaryl portion isoptionally substituted with one or more R⁵.

In one embodiment, compounds of formula (I) has the structure of formula(I″)

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is selected from a bond or C1-C3 alkylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—;

R¹, R², and R³ are each independently selected from phenyl, 5-10membered heteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl), 5-6membered heteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl),wherein each aryl or heteroaryl portion is optionally substituted withone or more R⁵;

R⁴ is each independently H or C1-C3 alkyl;

R⁵ is each independently I, Br, Cl, F, or C1-C3 alkyl; and

wherein the compound is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, a compound of formula (I), (I′), or (I″) is not

In one embodiment, the compound of formula (I) has the structure offormula (IA):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each phenyl, substituted with one or more R^(5a);

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5a) is each independently I, Br, Cl, F, C1-C6 alkyl, C1-C3 haloalkyl,—(C1-C6)-O—(C1-C6), C1-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, Oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (IB):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each phenyl, substituted with one or more R^(5a);

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5a) is each independently C1-C6 alkyl;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (IC):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each phenyl, substituted with one or more R^(5a), whereinat least one of R¹ and R² is,

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5a) is each independently I, Br, Cl, F, C1-C6 alkyl, C1-C3 haloalkyl,C1-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (ID):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (IE):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each —NHC(O)—;

R¹ and R² are each;

R³ is phenyl, optionally substituted with one or more R^(5b); and

R^(5b) is each independently I, Br, Cl, F, C1-C3 alkyl, C1-C3 haloalkyl,C1-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH.

In one embodiment, the compound of formula (I) has the structure offormula (IF):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each —NHC(O)—;

R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); and

R^(5b) is each independently C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy,C1-C3 haloalkoxy, OH, or COOH.

In one embodiment, the compound of formula (I) has the structure offormula (IG):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each —NHC(O)—;

R¹ and R² are each

R³ is phenyl; and

R^(5a) is each independently C1-C6 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy,C1-C3 haloalkoxy, OH, or COOH.

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or apharmaceutically acceptable excipient and a compound of any of formula(IA), (IB), (IC), (ID) (IE), (IF), and/or (IG). In one embodiment of thepresent disclosure, a method of modulating a Parkin ligase is providedcomprising administering to a subject in need thereof an effectiveamount of a compound of (IA), (IB), (IC), (ID) (IE), (IF), and/or (IG).

In one embodiment, the present disclosure provides compounds having thestructure of formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

M¹ and M² are each independently selected from a bond, —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, provided that M¹ and M² are not both —NR⁴— or both a bond;

R¹ and R² are each independently selected from a cycloalkyl, aryl,heterocyclyl, or heteroaryl, wherein each cycloalkyl, aryl, heteroaryl,and heterocyclyl is optionally substituted with one or more R^(5a),provided that R¹ and R² are not 1,3-dioxoisoindolin-2-yl;

R³ is selected from an alkyl, alkenyl, cycloalkyl, aryl, heterocyclyl,or heteroaryl, wherein each cycloalkyl, aryl, heteroaryl, andheterocyclyl is optionally substituted with one or more R^(5a);

R⁴ is each independently H or alkyl;

R^(5a) is each independently I, Br, Cl, F, CN, NH₂, NHR^(6a), NO₂,NR^(6a)R^(6a), OH, OR^(6a), or R^(6a); and

R^(6a) is each independently alkyl or haloalkyl; or alternatively twoR^(6a) on the same N atom can together form a 3-6 memberedN-heterocyclyl.

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or apharmaceutically acceptable excipient and a compound of Formula II. Inone embodiment of the present disclosure, a method of modulating aParkin ligase is provided comprising administering to a subject in needthereof an effective amount of a compound of Formula II.

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or apharmaceutically acceptable excipient and a compound of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L¹, L² and L³ are each independently selected from a bond, alkylene, oralkenylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, —NR⁴C(O)NR⁴—, —C(O)—, —C(═NR⁴)—, —C(═NOR⁴)—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, —NR⁴C(O)O—, —S(O)_(m)—, —S(O)_(m)NR⁴—, or—NR⁴S(O)_(m)—, provided that M¹ and M² are not both —NR⁴—;

R¹, R², and R³ are each independently selected from an alkyl,cycloalkyl, aryl, heterocycloalkyl, heterocyclyl, heteroaryl,cycloalkylalkyl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclylalkyl,heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl, wherein eachcycloalkyl, aryl, heteroaryl, and heterocyclyl portion is optionallysubstituted with one or more R⁵;

R⁴ is each independently H, alkyl, wherein each alkyl is optionallysubstituted with one or more R⁵;

R⁵ is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH,NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶,SO₃H, SO₃R⁶, or SR⁶;

R⁶ is each independently alkyl;

m is 0, 1, or 2; and

wherein the compound is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or apharmaceutically acceptable excipient and a compound of formula (I′):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³, M¹, M², R¹, R², and R³ are as defined for formula (I); and

wherein the compound is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or apharmaceutically acceptable excipient and a compound of formula (I″)

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is selected from a bond or C1-C3 alkylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—;

R¹, R², and R³ are each independently selected from phenyl, 5-10membered heteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl), 5-6membered heteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl),wherein each aryl or heteroaryl portion is optionally substituted withone or more R⁵;

R⁴ is each independently H or C1-C3 alkyl;

R⁵ is each independently I, Br, Cl, F, or C1-C3 alkyl; and

wherein the compound is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In another embodiment, the pharmaceutical composition comprising acompound of formula (I), (I′), or (I″), or a pharmaceutically acceptablesalt or solvate thereof further comprises one additional therapeuticallyactive agent.

In one embodiment of the present disclosure, a method of modulating aParkin ligase is provided comprising administering to a subject in needthereof an effective amount of a compound of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L¹, L² and L³ are each independently selected from a bond, alkylene, oralkenylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, —NR⁴C(O)NR⁴—, —C(O)—, —C(═NR⁴)—, —C(═NOR⁴)—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, —NR⁴C(O)O—, —S(O)_(m)—, —S(O)_(m)NR⁴—, or—NR⁴S(O)_(m)—, provided that M¹ and M² are not both —NR⁴—;

R¹, R², and R³ are each independently selected from an alkyl,cycloalkyl, aryl, heterocycloalkyl, heterocyclyl, heteroaryl,cycloalkylalkyl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclylalkyl,heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl, wherein eachcycloalkyl, aryl, heteroaryl, and heterocyclyl portion is optionallysubstituted with one or more R⁵;

R⁴ is each independently H, alkyl, wherein each alkyl is optionallysubstituted with one or more R⁵;

R⁵ is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH,NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶,SO₃H, SO₃R⁶, or SR⁶;

R⁶ is each independently alkyl; and

m is 0, 1, or 2.

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I), wherein L¹, L² and L³ are eachindependently selected from a bond, C1-C3 alkylene, or C2-C3 alkenylene.

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I), wherein M¹ and M² are eachindependently selected from —NR⁴—, —NR⁴C(O)—, —C(O)NR⁴—, NR⁴C(O)NR⁴—,—OC(O)—, —C(O)O—, —OC(O)O—, —OC(O)NR⁴—, or —NR⁴C(O)O—. In anotherembodiment, M¹ and M² are each independently selected from —NR⁴—,—NR⁴C(O)— or —C(O)NR⁴—.

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I), wherein R¹, R², and R³ are eachindependently selected from alkyl, aryl, heterocyclyl, heteroaryl,arylalkyl, arylalkenyl, heteroarylalkyl, heteroarylalkenyl, orheterocycloalkyl, wherein each cycloalkyl, aryl, heteroaryl andheterocyclyl portion is optionally substituted with one or more R⁵. Inone embodiment, R⁴ of formula (I) at each occurrence is independently Hor C1-C3 alkyl. In another embodiment, R¹, R², and R³ are eachindependently selected from C1-C3 alkyl, phenyl, 5-10 memberedheteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl), 5-6 memberedheteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl), wherein eachcycloalkyl, aryl, heteroaryl portion is optionally substituted with oneor more R⁵.

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I), wherein at least one of R¹, R²,and R³ is phenyl-(C2-C3 alkenyl). In another embodiment, at least one ofR¹, R², and R³ is a bicyclic 5-10 membered heteroaryl. In anotherembodiment, at least one of R¹, R², and R³ is

In another embodiment, at least one of R¹, R², and R³ is phenyl andsubstituted with at least one of methyl, I, Br, Cl or F.

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I), wherein L³ as a bond and R³ asan aryl. In another embodiment, L³ is a bond and R³ is a phenyl.

In one embodiment of the present disclosure, a method of modulating aParkin ligase is provided comprising administering to a subject in needthereof an effective amount of a compound of formula (I′):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³, M¹, M², R¹, R², and R³ are as defined for formula (I).

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I′), wherein M¹ and M² are eachindependently selected from —NR⁴—, —NR⁴C(O)—, —C(O)NR⁴—, NR⁴C(O)NR⁴—,—OC(O)—, —C(O)O—, —OC(O)O—, —OC(O)NR⁴—, or —NR⁴C(O)O—.

In one embodiment, the method disclosed herein comprises administeringto a subject a compound of formula (I′), wherein R¹, R², and R³ are eachindependently selected from alkyl, aryl, heteroaryl, arylalkyl,arylalkenyl, heteroarylalkyl, or heteroarylalkenyl, wherein eachcycloalkyl, aryl, heteroaryl portion is optionally substituted with oneor more R⁵.

In one embodiment of the present disclosure, a method of modulating aParkin ligase is provided comprising administering to a subject in needthereof an effective amount of a compound of formula (I″):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is selected from a bond or C1-C3 alkylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—;

R¹, R², and R³ are each independently selected from phenyl, 5-10membered heteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl), 5-6membered heteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl),wherein each aryl or heteroaryl portion is optionally substituted withone or more R⁵;

R⁴ is each independently H or C1-C3 alkyl; and

R⁵ is each independently I, Br, Cl, F, or C1-C3 alkyl.

In another embodiment of the present disclosure, a method of treating adisease or a condition is provided comprising administering to a subjectin need thereof a therapeutically effective amount of a compound offormula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L¹, L² and L³ are each independently selected from a bond, alkylene, oralkenylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, —NR⁴C(O)NR⁴—, —C(O)—, —C(═NR⁴)—, —C(═NOR⁴)—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, —NR⁴C(O)O—, —S(O)_(m)—, —S(O)_(m)NR⁴—, or—NR⁴S(O)_(m)—, provided that M¹ and M² are not both —NR⁴—;

R¹, R², and R³ are each independently selected from an alkyl,cycloalkyl, aryl, heterocycloalkyl, heterocyclyl, heteroaryl,cycloalkylalkyl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclylalkyl,heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl, wherein eachcycloalkyl, aryl, heteroaryl, and heterocyclyl portion is optionallysubstituted with one or more R⁵;

R⁴ is each independently H, alkyl, wherein each alkyl is optionallysubstituted with one or more R⁵;

R⁵ is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH,NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶,SO₃H, SO₃R⁶, or SR⁶;

R⁶ is each independently alkyl; and

m is 0, 1, or 2;

wherein the disease or the condition is selected from the groupconsisting of cancer, neurological disease, a disorder characterized byabnormal accumulation of α-synuclein, a disorder of an aging process,cardiovascular disease, bacterial infection, viral infection,mitochondrial related disease, mental retardation, deafness, blindness,diabetes, obesity, autoimmune disease, glaucoma, Leber's HereditaryOptic Neuropathy, and rheumatoid arthritis.

In another embodiment of the present disclosure, a method of treating adisease or a condition is provided comprising administering to a subjectin need thereof a therapeutically effective amount of a compound offormula (I′):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³, M¹, M², R¹, R², and R³ are as defined for formula (I);

wherein the disease or the condition is selected from the groupconsisting of cancer, neurological disease, a disorder characterized byabnormal accumulation of α-synuclein, a disorder of an aging process,cardiovascular disease, bacterial infection, viral infection,mitochondrial related disease, mental retardation, deafness, blindness,diabetes, obesity, autoimmune disease, glaucoma, Leber's HereditaryOptic Neuropathy, and rheumatoid arthritis.

In another embodiment of the present disclosure, a method of treating adisease or a condition is provided comprising administering to a subjectin need thereof a therapeutically effective amount of a compound offormula (I″):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is selected from a bond or C1-C3 alkylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—;

R¹, R², and R³ are each independently selected from phenyl, 5-10membered heteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl), 5-6membered heteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl),wherein each aryl or heteroaryl portion is optionally substituted withone or more R⁵;

R⁴ is each independently H or C1-C3 alkyl; and

R⁵ is each independently I, Br, Cl, F, or C1-C3 alkyl;

wherein the disease or the condition is selected from the groupconsisting of cancer, neurological disease, a disorder characterized byabnormal accumulation of α-synuclein, a disorder of an aging process,cardiovascular disease, bacterial infection, viral infection,mitochondrial related disease, mental retardation, deafness, blindness,diabetes, obesity, autoimmune disease, glaucoma, Leber's HereditaryOptic Neuropathy, and rheumatoid arthritis.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 indicates thatN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide (Compound A)increases the Parkin Ligase reaction with the Activity-based Ubiquitinvinyl sulfone probe.

FIG. 2 indicates that compoundN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl) (Compound A) dibenzamideincreases Parkin activity in an auto-ubiquitination assay.

FIG. 3 shows mitophagy cell assay result forN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide (Compound A).

FIG. 4 indicates thatN-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide (Compound C)increases the Parkin Ligase reaction with the Activity-based Ubiquitinvinyl sulfone probe.

FIG. 5 indicates that compoundN-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide (Compound C)increases Parkin activity in an auto-ubiquitination assay.

FIG. 6 shows mitophagy cell assay result forN-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide (Compound C).

FIG. 7 shows rat intravenous (IV) and oral (PO) bioavailability forCompound A.

FIG. 8 shows rat intravenous (IV) and intraperitoneal (PO)bioavailability for Compound F.

FIG. 9 shows rat intravenous (IV) and oral (PO) bioavailability forCompound K.

FIG. 10 shows rat intravenous (IV) and oral (PO) bioavailability forCompound H.

FIG. 11 shows rat intravenous (IV) and oral (PO) bioavailability forCompound C.

FIG. 12 shows a Xenograft study testing Compound F efficacy to delaysubcutaneous HCT-116 tumor growth. Compound F or controls wereadministered daily (IP). Group 1=Vehicle Control, Group 2=Bevacizumab (5mg/kg), Group 3=Compound F (5 mg/kg), Group 4=Compound F (10 mg/kg)

FIG. 13 shows a Xenograft study testing Compound F efficacy to delaysubcutaneous Calu-6 tumor growth. Compound F or controls wereadministered daily (IP). Group 1=Vehicle Control, Group 2=Bevacizumab (5mg/kg), Group 3=Compound F (5 mg/kg), Group 4=Compound F (10 mg/kg).

FIGS. 14A-14K shows the % inhibition of cancer cell line proliferationfrom various compound candidates.

DETAILED DESCRIPTION

All publications, patents and patent applications, including anydrawings and appendices therein are incorporated by reference in theirentirety for all purposes to the same extent as if each individualpublication, patent or patent application, drawing, or appendix wasspecifically and individually indicated to be incorporated by referencein its entirety for all purposes.

Definitions

While the following terms are believed to be well understood by one ofordinary skill in the art, the following definitions are set forth tofacilitate explanation of the presently disclosed subject matter.

Throughout the present specification, the terms “about” and/or“approximately” may be used in conjunction with numerical values and/orranges. The term “about” is understood to mean those values near to arecited value. For example, “about 40 [units]” may mean within ±25% of40 (e.g., from 30 to 50), within ±20%, ±15%, ±10%, ±9%, ±8%, ±7%, ±6%,±5%, ±4%, ±3%, ±2%, ±1%, less than ±1%, or any other value or range ofvalues therein or therebelow. Furthermore, the phrases “less than about[a value]” or “greater than about [a value]” should be understood inview of the definition of the term “about” provided herein. The terms“about” and “approximately” may be used interchangeably.

Throughout the present specification, numerical ranges are provided forcertain quantities. It is to be understood that these ranges compriseall subranges therein. Thus, the range “from 50 to 80” includes allpossible ranges therein (e.g., 51-79, 52-78, 53-77, 54-76, 55-75, 60-70,etc.). Furthermore, all values within a given range may be an endpointfor the range encompassed thereby (e.g., the range 50-80 includes theranges with endpoints such as 55-80, 50-75, etc.).

The term “a” or “an” refers to one or more of that entity; for example,“a kinase inhibitor” refers to one or more kinase inhibitors or at leastone kinase inhibitor. As such, the terms “a” (or “an”), “one or more”and “at least one” are used interchangeably herein. In addition,reference to “an inhibitor” by the indefinite article “a” or “an” doesnot exclude the possibility that more than one of the inhibitors ispresent, unless the context clearly requires that there is one and onlyone of the inhibitors.

As used herein, the verb “comprise” as is used in this description andin the claims and its conjugations are used in its non-limiting sense tomean that items following the word are included, but items notspecifically mentioned are not excluded. The present invention maysuitably “comprise”, “consist of”, or “consist essentially of”, thesteps, elements, and/or reagents described in the claims.

It is further noted that the claims may be drafted to exclude anyoptional element. As such, this statement is intended to serve asantecedent basis for use of such exclusive terminology as “solely”,“only” and the like in connection with the recitation of claim elements,or the use of a “negative” limitation.

The term “pharmaceutically acceptable salts” include those obtained byreacting the active compound functioning as a base, with an inorganic ororganic acid to form a salt, for example, salts of hydrochloric acid,sulfuric acid, phosphoric acid, methanesulfonic acid, camphorsulfonicacid, oxalic acid, maleic acid, succinic acid, citric acid, formic acid,hydrobromic acid, benzoic acid, tartaric acid, fumaric acid, salicylicacid, mandelic acid, carbonic acid, etc. Those skilled in the art willfurther recognize that acid addition salts may be prepared by reactionof the compounds with the appropriate inorganic or organic acid via anyof a number of known methods.

The term “treating” means one or more of relieving, alleviating,delaying, reducing, reversing, improving, or managing at least onesymptom of a condition in a subject. The term “treating” may also meanone or more of arresting, delaying the onset (i.e., the period prior toclinical manifestation of the condition) or reducing the risk ofdeveloping or worsening a condition.

An “effective amount” means the amount of a formulation according to theinvention that, when administered to a patient for treating a state,disorder or condition is sufficient to effect such treatment. The“effective amount” will vary depending on the active ingredient, thestate, disorder, or condition to be treated and its severity, and theage, weight, physical condition and responsiveness of the mammal to betreated.

The term “therapeutically effective” applied to dose or amount refers tothat quantity of a compound or pharmaceutical formulation that issufficient to result in a desired clinical benefit after administrationto a patient in need thereof.

All weight percentages (i.e., “% by weight” and “wt. %” and w/w)referenced herein, unless otherwise indicated, are measured relative tothe total weight of the pharmaceutical composition.

As used herein, “substantially” or “substantial” refers to the completeor nearly complete extent or degree of an action, characteristic,property, state, structure, item, or result. For example, an object thatis “substantially” enclosed would mean that the object is eithercompletely enclosed or nearly completely enclosed. The exact allowabledegree of deviation from absolute completeness may in some cases dependon the specific context. However, generally speaking, the nearness ofcompletion will be so as to have the same overall result as if absoluteand total completion were obtained. The use of “substantially” isequally applicable when used in a negative connotation to refer to thecomplete or near complete lack of action, characteristic, property,state, structure, item, or result. For example, a composition that is“substantially free of” other active agents would either completely lackother active agents, or so nearly completely lack other active agentsthat the effect would be the same as if it completely lacked otheractive agents. In other words, a composition that is “substantially freeof” an ingredient or element or another active agent may still containsuch an item as long as there is no measurable effect thereof

As used herein, the “alignment” of two or more protein/amino acidsequences may be performed using the alignment program ClustalW2,available at www.ebi.ac.uk/Tools/msa/clustalw2/. The following defaultparameters may be used for Pairwise alignment: Protein WeightMatrix=Gonnet; Gap Open=10; Gap Extension=0.1.

“Ubiquitin Proteasome Pathway System (UPS)” as used herein relates tothe ubiquitin proteasome pathway, conserved from yeast to mammals, andis required for the targeted degradation of most short-lived proteins inthe eukaryotic cell. Targets include cell cycle regulatory proteins,whose timely destruction is vital for controlled cell division, as wellas proteins unable to fold properly within the endoplasmic reticulum.Ubiquitin modification is an ATP-dependent process carried out by threeclasses of enzymes. An “ubiquitin activating enzyme” (E1) forms athio-ester bond with ubiquitin, a highly conserved 76-amino acidprotein. This reaction allows subsequent binding of ubiquitin to a“ubiquitin conjugating enzyme” (E2), followed by the formation of anisopeptide bond between the carboxy-terminus of ubiquitin and a lysineresidue on the substrate protein. The latter reaction requires a“ubiquitin ligase” (E3). E3 ligases can be single- or multi-subunitenzymes. In some cases, the ubiquitin-binding and substrate bindingdomains reside on separate polypeptides brought together by adaptorproteins or culling. Numerous E3 ligases provide specificity in thateach can modify only a subset of substrate proteins. Further specificityis achieved by post-translational modification of substrate proteins,including, but not limited to, phosphorylation. Effects ofmonoubiquitination include changes in subcellular localization. However,multiple ubiquitination cycles resulting in a polyubiquitin chain arerequired for targeting a protein to the proteasome for degradation. Themultisubunit 26S proteasome recognizes, unfolds, and degradespolyubiquitinated substrates into small peptides. The reaction occurswithin the cylindrical core of the proteasome complex, and peptide bondhydrolysis employs a core threonine residue as the catalyticnucleophile. It has been shown that an additional layer of complexity,in the form of multiubiquitin chain receptors, may lie between thepolyubiquitination and degradation steps. These receptors react with asubset of polyubiquitinated substrates, aiding in their recognition bythe 26S proteasome, and thereby promoting their degradation. Thispathway is not only important in cellular homeostasis, but also in humandisease. Because ubiquitin/proteasome-dependent degradation is oftenemployed in control of the cell division cycle and cell growth,researchers have found that proteasome inhibitors hold some promise ofbeing developed into potential cancer therapeutic agents.

Protein degradation through the ubiquitin-proteasome system is the majorpathway of non-lysosomal proteolysis of intracellular proteins. It playsimportant roles in a variety of fundamental cellular processes such asregulation of cell cycle progression, division, development anddifferentiation, apoptosis, cell trafficking, and modulation of theimmune and inflammatory responses. The central element of this system isthe covalent linkage of ubiquitin to targeted proteins, which are thenrecognized by the 26S proteasome, an adenosine triphosphate-dependent,multi-catalytic protease. Damaged, oxidized, or misfolded proteins aswell as regulatory proteins that control many critical cellularfunctions are among the targets of this degradation process. Aberrationof this system leads to the dysregulation of cellular homeostasis andthe development of multiple diseases (Wang et al. Cell Mol Immunol. 2006August; 3(4):255-61).

“Parkin ligase” or “Parkin” as used herein relates to a protein which inhumans is encoded by the PARK2 gene. (Kitada T, Asakawa S, Hattori N,Matsumine H, Yamamura Y, Minoshima S, Yokochi M, Mizuno Y, Shimizu N(April 1998). “Mutations in the parkin gene cause autosomal recessivejuvenile parkinsonism”. Nature 392 (6676): 605-608. doi:10.1038/33416.PMID 9560156. Matsumine H, Yamamura Y, Hattori N, Kobayashi T, Kitada T,Yoritaka A, Mizuno Y (April 1998). “A microdeletion of D6S305 in afamily of autosomal recessive juvenile parkinsonism (PARK2)”. Genomics49 (1): 143-146. doi:10.1006/geno.1997.5196. PMID 9570960. The proteinis a component of a multiprotein E3 ubiquitin ligase complex which inturn is part of the ubiquitin-proteasome system that mediates thetargeting of proteins for degradation. Mutations in the PARK2 gene areknown to cause a familial form of Parkinson's disease known as autosomalrecessive juvenile Parkinson's disease (AR-JP).

“Ligase” as used herein, is an enzyme that can catalyze the joining oftwo or more compounds or biomolecules by bonding them together with anew chemical bond. The “ligation” of the two usually with accompanyinghydrolysis of a small chemical group dependent to one of the largercompounds or biomolecules, or the enzyme catalyzing the linking togetherof two compounds, e.g., enzymes that catalyze joining of groups C—O,C—S, C—N, etc. Ubiquitin-protein (E3) ligases are a large family ofhighly diverse enzymes selecting proteins for ubiquitination.

“Ub Ligases” are involved in disease pathogenesis for oncology,inflammation & infectious disease. E3 ligase belonging to theRING-between-RING (RBR) family of E3 ligases containing both canonicalRING domains and a catalytic cysteine residue usually restricted to HECTE3 ligases; termed ‘RING/HECT hybrid’ enzymes. Mutations in Parkinlinked to Parkinson's disease, cancer and mycobacterial infection.Parkin is recognized as a neuroprotective protein with a role inmitochondrial integrity. Human genetic data implicate loss of Parkinactivity as a mechanism for pathogenesis of Parkinson's disease (PD).

“Zinc Finger (ZnF) Domain” as used herein relates to a protein structurecharacterized by coordinating zinc ions to stabilize the functionalactivity. ZnF stabilize the binding of Ub, Deubiquitinating Enzymes(DUBs), and Ligases (E3) in the UPS.

“Ligands” as used herein bind to metal via one or more atoms in theligand, and are often termed as chelating ligands. A ligand that bindsthrough two sites is classified as bidentate, and three sites astridentate. The “bite angle” refers to the angle between the two bondsof a bidentate chelate. Chelating ligands are commonly formed by linkingdonor groups via organic linkers. A classic bidentate ligand isethylenediamine, which is derived by the linking of two ammonia groupswith an ethylene (—CH2CH2-) linker. A classic example of a polydentateligand is the hexadentate chelating agent EDTA, which is able to bondthrough six sites, completely surrounding some metals. The bindingaffinity of a chelating system depends on the chelating angle or biteangle. Many ligands are capable of binding metal ions through multiplesites, usually because the ligands have lone pairs on more than oneatom. Some ligands can bond to a metal center through the same atom butwith a different number of lone pairs. The bond order of the metalligand bond can be in part distinguished through the metal ligand bondangle (M-X-R). This bond angle is often referred to as being linear orbent with further discussion concerning the degree to which the angle isbent. For example, an imido ligand in the ionic form has three lonepairs. One lone pair is used as a sigma X donor, the other two lonepairs are available as L type pi donors. If both lone pairs are used inpi bonds then the M-N-R geometry is linear. However, if one or both ofthese lone pairs are non-bonding then the M-N-R bond is bent and theextent of the bend speaks to how much pi bonding there may be. It wasfound that few heteroatoms, such as nitrogen, oxygen, and sulfur atoms,interacted with zinc, ideal distances between the zinc and theseheteroatoms were identified. Whereas carboxylates bound to the zinc viaboth monodentate and bidentate interactions, the hydroxamates bounddominantly in a bidentate manner. These results aid in the design of newinhibitors with the potential to interact with zinc in the targetprotein. Virtually every molecule and every ion can serve as a ligandfor (or “coordinate to”) metals. Monodentate ligands include virtuallyall anions and all simple Lewis bases. Thus, the halides andpseudohalides are important anionic ligands whereas ammonia, carbonmonoxide, and water are particularly common charge-neutral ligands.Simple organic species are also very common, be they anionic (RO⁻ andRCO₂ ⁻) or neutral (R₂O, R₂S, R_(3-x)NH_(x), and R₃P). Complexes ofpolydentate ligands are called chelate complexes. They tend to be morestable than complexes derived from monodentate ligands. This enhancedstability, the chelate effect, is usually attributed to effects ofentropy, which favors the displacement of many ligands by onepolydentate ligand. When the chelating ligand forms a large ring that atleast partially surrounds the central atom and bonds to it, leaving thecentral atom at the center of a large ring. The more rigid and thehigher its denticity, the more inert will be the macrocyclic complex.

“Chelator” as used herein relates to a binding agent that suppresseschemical activity by forming a chelate (a coordination compound in whicha metal atom or ion is bound to a ligand at two or more points on theligand, so as to form, for example, a heterocyclic ring containing ametal atom).

“Chelation” as used herein relates to a particular way that ions andmolecules bind metal ions. According to the International Union of Pureand Applied Chemistry (IUPAC), chelation involves the formation orpresence of two or more separate coordinate bonds between a polydentate(multiple bonded) ligand and a single central atom. Usually theseligands are organic compounds, and are called chelants, chelators,chelating agents, or sequestering agents.

“Electrophile” as used herein relates to species that is attracted to anelectron rich center. In chemistry, an electrophile is a reagentattracted to electrons. It participates in a chemical reaction byaccepting an electron pair in order to bond to a nucleophile. Becauseelectrophiles accept electrons, they are Lewis acids. Most electrophilesare positively charged, have an atom that carries a partial positivecharge, or have an atom that does not have an octet of electrons.

The terms below, as used herein, have the following meanings, unlessindicated otherwise:

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodo radical.

“Hydroxy” or “hydroxyl” refers to the —OH radical.

“Imino” refers to the ═NH substituent.

“Nitro” refers to the —NO₂ radical.

“Oxo” refers to the ═O substituent.

“Thioxo” refers to the ═S substituent.

“Alkyl” or “alkyl group” refers to a fully saturated, straight orbranched hydrocarbon chain radical having from one to twelve carbonatoms, and which is attached to the rest of the molecule by a singlebond. Alkyls comprising any number of carbon atoms from 1 to 12 areincluded. An alkyl comprising up to 12 carbon atoms is a C₁-C₁₂ alkyl,an alkyl comprising up to 10 carbon atoms is a C₁-C₁₀ alkyl, an alkylcomprising up to 6 carbon atoms is a C₁-C₆ alkyl and an alkyl comprisingup to 5 carbon atoms is a C₁-C₅ alkyl. A C₁-C₅ alkyl includes C₅ alkyls,C₄ alkyls, C₃ alkyls, C₂ alkyls and C₁ alkyl (i.e., methyl). A C₁-C₆alkyl includes all moieties described above for C₁-C₅ alkyls but alsoincludes C₆ alkyls. A C₁-C₁₀ alkyl includes all moieties described abovefor C₁-C₅ alkyls and C₁-C₆ alkyls, but also includes C₇, C₈, C₉ and C₁₀alkyls. Similarly, a C₁-C₁₂ alkyl includes all the foregoing moieties,but also includes C₁₁ and C₁₂ alkyls. Non-limiting examples of C₁-C₁₂alkyl include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl,i-butyl, sec-butyl, t-butyl, n-pentyl, t-amyl, n-hexyl, n-heptyl,n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless statedotherwise specifically in the specification, an alkyl group can beoptionally substituted.

“Alkylene” or “alkylene chain” refers to a fully saturated, straight orbranched divalent hydrocarbon chain radical, and having from one totwelve carbon atoms. Non-limiting examples of C₁-C₁₂ alkylene includemethylene, ethylene, propylene, n-butylene, ethenylene, propenylene,n-butenylene, propynylene, n-butynylene, and the like. The alkylenechain is attached to the rest of the molecule through a single bond andto the radical group through a single bond. The points of attachment ofthe alkylene chain to the rest of the molecule and to the radical groupcan be through one carbon or any two carbons within the chain. Unlessstated otherwise specifically in the specification, an alkylene chaincan be optionally substituted.

“Alkenyl” or “alkenyl group” refers to a straight or branchedhydrocarbon chain radical having from two to twelve carbon atoms, andhaving one or more carbon-carbon double bonds. Each alkenyl group isattached to the rest of the molecule by a single bond. Alkenyl groupcomprising any number of carbon atoms from 2 to 12 are included. Analkenyl group comprising up to 12 carbon atoms is a C₂-C₁₂ alkenyl, analkenyl comprising up to 10 carbon atoms is a C₂-C₁₀ alkenyl, an alkenylgroup comprising up to 6 carbon atoms is a C₂-C₆ alkenyl and an alkenylcomprising up to 5 carbon atoms is a C₂-C₅ alkenyl. A C₂-C₅ alkenylincludes C₅ alkenyls, C₄ alkenyls, C₃ alkenyls, and C₂ alkenyls. A C₂-C₆alkenyl includes all moieties described above for C₂-C₅ alkenyls butalso includes C₆ alkenyls. A C₂-C₁₀ alkenyl includes all moietiesdescribed above for C₂-C₅ alkenyls and C₂-C₆ alkenyls, but also includesC₇, C₈, C₉ and C₁₀ alkenyls. Similarly, a C₂-C₁₂ alkenyl includes allthe foregoing moieties, but also includes C₁₁ and C₁₂ alkenyls.Non-limiting examples of C₂-C₁₂ alkenyl include ethenyl (vinyl),1-propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-1-propenyl,1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl,4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl,1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl,1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl,7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 4-nonenyl, 5-nonenyl,6-nonenyl, 7-nonenyl, 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl,4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl,1-undecenyl, 2-undecenyl, 3-undecenyl, 4-undecenyl, 5-undecenyl,6-undecenyl, 7-undecenyl, 8-undecenyl, 9-undecenyl, 10-undecenyl,1-dodecenyl, 2-dodecenyl, 3-dodecenyl, 4-dodecenyl, 5-dodecenyl,6-dodecenyl, 7-dodecenyl, 8-dodecenyl, 9-dodecenyl, 10-dodecenyl, and11-dodecenyl. Unless stated otherwise specifically in the specification,an alkyl group can be optionally substituted.

“Alkenylene” or “alkenylene chain” refers to a straight or brancheddivalent hydrocarbon chain radical, having from two to twelve carbonatoms, and having one or more carbon-carbon double bonds. Non-limitingexamples of C₂-C₁₂ alkenylene include ethene, propene, butene, and thelike. The alkenylene chain is attached to the rest of the moleculethrough a single bond and to the radical group through a single bond.The points of attachment of the alkenylene chain to the rest of themolecule and to the radical group can be through one carbon or any twocarbons within the chain. Unless stated otherwise specifically in thespecification, an alkenylene chain can be optionally substituted.

“Alkynyl” or “alkynyl group” refers to a straight or branchedhydrocarbon chain radical having from two to twelve carbon atoms, andhaving one or more carbon-carbon triple bonds. Each alkynyl group isattached to the rest of the molecule by a single bond. Alkynyl groupcomprising any number of carbon atoms from 2 to 12 are included. Analkynyl group comprising up to 12 carbon atoms is a C₂-C₁₂ alkynyl, analkynyl comprising up to 10 carbon atoms is a C₂-C10 alkynyl, an alkynylgroup comprising up to 6 carbon atoms is a C₂-C₆ alkynyl and an alkynylcomprising up to 5 carbon atoms is a C₂-C₅ alkynyl. A C₂-C₅ alkynylincludes C₅ alkynyls, C₄ alkynyls, C₃ alkynyls, and C₂ alkynyls. A C₂-C₆alkynyl includes all moieties described above for C₂-C₅ alkynyls butalso includes C₆ alkynyls. A C₂-C₁₀ alkynyl includes all moietiesdescribed above for C2-C₅ alkynyls and C₂-C₆ alkynyls, but also includesC₇, C₈, C₉ and C₁₀ alkynyls. Similarly, a C₂-C₁₂ alkynyl includes allthe foregoing moieties, but also includes C₁₁ and C₁₂ alkynyls.Non-limiting examples of C₂-C₁₂ alkenyl include ethynyl, propynyl,butynyl, pentynyl and the like. Unless stated otherwise specifically inthe specification, an alkyl group can be optionally substituted.

“Alkynylene” or “alkynylene chain” refers to a straight or brancheddivalent hydrocarbon chain radical, having from two to twelve carbonatoms, and having one or more carbon-carbon triple bonds. Non-limitingexamples of C₂-C₁₂ alkynylene include ethynylene, propargylene and thelike. The alkynylene chain is attached to the rest of the moleculethrough a single bond and to the radical group through a single bond.The points of attachment of the alkynylene chain to the rest of themolecule and to the radical group can be through one carbon or any twocarbons within the chain. Unless stated otherwise specifically in thespecification, an alkynylene chain can be optionally substituted.

“Alkoxy” refers to a radical of the formula —OR_(a) where R_(a) is analkyl, alkenyl or alknyl radical as defined above containing one totwelve carbon atoms. Unless stated otherwise specifically in thespecification, an alkoxy group can be optionally substituted.

“Alkylamino” refers to a radical of the formula —NHR_(a) or —NR_(a)R_(a)where each R_(a) is, independently, an alkyl, alkenyl or alkynyl radicalas defined above containing one to twelve carbon atoms. Unless statedotherwise specifically in the specification, an alkylamino group can beoptionally substituted.

“Alkylcarbonyl” refers to the —C(═O)R_(a) moiety, wherein R_(a) is analkyl, alkenyl or alkynyl radical as defined above. A non-limitingexample of an alkyl carbonyl is the methyl carbonyl (“acetal”) moiety.Alkylcarbonyl groups can also be referred to as “Cw-Cz acyl” where w andz depicts the range of the number of carbon in R_(a), as defined above.For example, “C₁-C₁₀ acyl” refers to alkylcarbonyl group as definedabove, where R_(a) is C₁-C₁₀ alkyl, C₁-C₁₀ alkenyl, or C₁-C₁₀ alkynylradical as defined above. Unless stated otherwise specifically in thespecification, an alkyl carbonyl group can be optionally substituted.

“Aryl” refers to a hydrocarbon ring system radical comprising hydrogen,6 to 18 carbon atoms and at least one aromatic ring. For purposes ofthis invention, the aryl radical can be a monocyclic, bicyclic,tricyclic or tetracyclic ring system, which can include fused or bridgedring systems. Aryl radicals include, but are not limited to, arylradicals derived from aceanthrylene, acenaphthylene, acephenanthrylene,anthracene, azulene, benzene, chrysene, fluoranthene, fluorene,as-indacene, s-indacene, indane, indene, naphthalene, phenalene,phenanthrene, pleiadene, pyrene, and triphenylene. Unless statedotherwise specifically in the specification, the term “aryl” is meant toinclude aryl radicals that are optionally substituted.

“Aralkyl” or “arylalkyl” refers to a radical of the formula —R_(b)—R_(c)where R_(b) is an alkylene group as defined above and R_(c) is one ormore aryl radicals as defined above, for example, benzyl, diphenylmethyland the like. Unless stated otherwise specifically in the specification,an aralkyl group can be optionally substituted.

“Aralkenyl” or “arylalkenyl” refers to a radical of the formula—R_(b)—R_(c) where R_(b) is an alkenylene o group as defined above andR_(c) is one or more aryl radicals as defined above. Unless statedotherwise specifically in the specification, an aralkenyl group can beoptionally substituted.

“Aralkynyl” or “arylalkynyl” refers to a radical of the formula—R_(b)—R_(c) where R_(b) is an alkynylene group as defined above andR_(c) is one or more aryl radicals as defined above. Unless statedotherwise specifically in the specification, an aralkynyl group can beoptionally substituted.

“Carbocyclyl,” “carbocyclic ring” or “carbocycle” refers to a ringsstructure, wherein the atoms which form the ring are each carbon.Carbocyclic rings can comprise from 3 to 20 carbon atoms in the ring.Carbocyclic rings include aryls and cycloalkyl. Cycloalkenyl andcycloalkynyl as defined herein. Unless stated otherwise specifically inthe specification, a carbocyclyl group can be optionally substituted.

“Cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclicfully saturated hydrocarbon radical consisting solely of carbon andhydrogen atoms, which can include fused or bridged ring systems, havingfrom three to twenty carbon atoms, preferably having from three to tencarbon atoms, and which is attached to the rest of the molecule by asingle bond. Monocyclic cycloalkyl radicals include, for example,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, andcyclooctyl. Polycyclic cycloalkyl radicals include, for example,adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl,and the like. Unless otherwise stated specifically in the specification,a cycloalkyl group can be optionally substituted.

“Cycloalkenyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,having one or more carbon-carbon double bonds, which can include fusedor bridged ring systems, having from three to twenty carbon atoms,preferably having from three to ten carbon atoms, and which is attachedto the rest of the molecule by a single bond. Monocyclic cycloalkenylradicals include, for example, cyclopentenyl, cyclohexenyl,cycloheptenyl, cycloctenyl, and the like. Polycyclic cycloalkenylradicals include, for example, bicyclo[2.2.1]hept-2-enyl and the like.Unless otherwise stated specifically in the specification, acycloalkenyl group can be optionally substituted.

“Cycloalkynyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,having one or more carbon-carbon triple bonds, which can include fusedor bridged ring systems, having from three to twenty carbon atoms,preferably having from three to ten carbon atoms, and which is attachedto the rest of the molecule by a single bond. Monocyclic cycloalkynylradicals include, for example, cycloheptynyl, cyclooctynyl, and thelike. Unless otherwise stated specifically in the specification, acycloalkynyl group can be optionally substituted.

“Cycloalkylalkyl” refers to a radical of the formula —R_(b)—R_(d) whereR_(b) is an alkylene, alkenylene, or alkynylene group as defined aboveand R_(d) is a cycloalkyl, cycloalkenyl, cycloalkynyl radical as definedabove. Unless stated otherwise specifically in the specification, acycloalkylalkyl group can be optionally substituted.

“Haloalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl,1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and thelike. Unless stated otherwise specifically in the specification, ahaloalkyl group can be optionally substituted.

“Haloalkenyl” refers to an alkenyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,1-fluoropropenyl, 1,1-difluorobutenyl, and the like. Unless statedotherwise specifically in the specification, a haloalkenyl group can beoptionally substituted.

“Haloalkynyl” refers to an alkynyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,1-fluoropropynyl, 1-fluorobutynyl, and the like. Unless stated otherwisespecifically in the specification, a haloalkenyl group can be optionallysubstituted.

“Heterocyclyl,” “heterocyclic ring” or “heterocycle” refers to a stable3- to 20-membered non-aromatic, partially aromatic, or aromatic ringradical which consists of two to twelve carbon atoms and from one to sixheteroatoms selected from the group consisting of nitrogen, oxygen andsulfur. Heterocyclycl or heterocyclic rings include heteroaryls asdefined below. Unless stated otherwise specifically in thespecification, the heterocyclyl radical can be a monocyclic, bicyclic,tricyclic or tetracyclic ring system, which can include fused or bridgedring systems; and the nitrogen, carbon or sulfur atoms in theheterocyclyl radical can be optionally oxidized; the nitrogen atom canbe optionally quaternized; and the heterocyclyl radical can be partiallyor fully saturated. Examples of such heterocyclyl radicals include, butare not limited to, dioxolanyl, thienyl[1,3]dithianyl,decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl,isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl,2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl,piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl,quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl,tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl,1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless statedotherwise specifically in the specification, a heterocyclyl group can beoptionally substituted.

“Heterocyclylalkyl” refers to a radical of the formula —R_(b)—R_(e)where R_(b) is an alkylene group as defined above and R_(e) is aheterocyclyl radical as defined above. Unless stated otherwisespecifically in the specification, a heterocycloalkylalkyl group can beoptionally substituted.

“Heterocyclylalkenyl” refers to a radical of the formula —R_(b)—R_(e)where R_(b) is an alkenylene group as defined above and R_(e) is aheterocyclyl radical as defined above. Unless stated otherwisespecifically in the specification, a heterocycloalkylalkenyl group canbe optionally substituted.

“Heterocyclylalkynyl” refers to a radical of the formula —R_(b)—R_(e)where R_(b) is an alkynylene group as defined above and R_(e) is aheterocyclyl radical as defined above. Unless stated otherwisespecifically in the specification, a heterocycloalkylalkynyl group canbe optionally substituted.

“N-heterocyclyl” refers to a heterocyclyl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heterocyclyl radical to the rest of the molecule is through anitrogen atom in the heterocyclyl radical. Unless stated otherwisespecifically in the specification, a N-heterocyclyl group can beoptionally substituted.

“Heteroaryl” refers to a 5- to 20-membered ring system radicalcomprising hydrogen atoms, one to thirteen carbon atoms, one to sixheteroatoms selected from the group consisting of nitrogen, oxygen andsulfur, and at least one aromatic ring. For purposes of this invention,the heteroaryl radical can be a monocyclic, bicyclic, tricyclic ortetracyclic ring system, which can include fused or bridged ringsystems; and the nitrogen, carbon or sulfur atoms in the heteroarylradical can be optionally oxidized; the nitrogen atom can be optionallyquaternized. Examples include, but are not limited to, azepinyl,acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl,benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl,benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl(benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl,carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl,furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl,isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl,isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl,1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl,phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl,pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl,quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl,tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl,triazinyl, and thiophenyl (i.e. thienyl). Unless stated otherwisespecifically in the specification, a heteroaryl group can be optionallysubstituted.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. Unless stated otherwise specifically inthe specification, an N-heteroaryl group can be optionally substituted.

“Heteroarylalkyl” refers to a radical of the formula —R_(b)—R_(f) whereR_(b) is an alkylene chain as defined above and R_(f) is a heteroarylradical as defined above. Unless stated otherwise specifically in thespecification, a heteroarylalkyl group can be optionally substituted.

“Heteroarylalkenyl” refers to a radical of the formula —R_(b)—R_(f)where R_(b) is an alkenylene, chain as defined above and R_(f) is aheteroaryl radical as defined above. Unless stated otherwisespecifically in the specification, a heteroarylalkenyl group can beoptionally substituted.

“Heteroarylalkynyl” refers to a radical of the formula —R_(b)—R_(f)where R_(b) is an alkynylene chain as defined above and R_(f) is aheteroaryl radical as defined above. Unless stated otherwisespecifically in the specification, a heteroarylalkynyl group can beoptionally substituted.

“Thioalkyl” refers to a radical of the formula —SR_(a) where R_(a) is analkyl, alkenyl, or alkynyl radical as defined above containing one totwelve carbon atoms. Unless stated otherwise specifically in thespecification, a thioalkyl group can be optionally substituted.

The term “substituted” used herein means any of the above groups (i.e.,alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, alkoxy,alkylamino, alkylcarbonyl, thioalkyl, aryl, aralkyl, carbocyclyl,cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl,heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl,N-heteroaryl and/or heteroarylalkyl) wherein at least one hydrogen atomis replaced by a bond to a non-hydrogen atoms such as, but not limitedto: a halogen atom such as F, Cl, Br, and I; an oxygen atom in groupssuch as hydroxyl groups, alkoxy groups, and ester groups; a sulfur atomin groups such as thiol groups, thioalkyl groups, sulfone groups,sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such asamines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines,diarylamines, N-oxides, imides, and enamines; a silicon atom in groupssuch as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilylgroups, and triarylsilyl groups; and other heteroatoms in various othergroups. “Substituted” also means any of the above groups in which one ormore hydrogen atoms are replaced by a higher-order bond (e.g., a double-or triple-bond) to a heteroatom such as oxygen in oxo, carbonyl,carboxyl, and ester groups; and nitrogen in groups such as imines,oximes, hydrazones, and nitriles. For example, “substituted” includesany of the above groups in which one or more hydrogen atoms are replacedwith —NR_(g)R_(h), —NR_(g)C(═O)R_(h), —NR_(g)C(═O)NR_(g)R_(h),—NR_(g)C(═O)OR_(h), —NR_(g)SO₂R_(h), —OC(═O)NR_(g)R_(h), —OR_(g),—SR_(g), —SOR_(g), —SO₂R_(g), —OSO₂R_(g), —SO₂OR_(g), ═NSO₂R_(g), and—SO₂NR_(g)R_(h). “Substituted” also means any of the above groups inwhich one or more hydrogen atoms are replaced with —C(═O)R_(g),—C(═O)OR_(g), —C(═O)NR_(g)R_(h), —CH₂SO₂R_(g), —CH₂SO₂NR_(g)R_(h). Inthe foregoing, R_(g) and R_(h) are the same or different andindependently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino,thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl,cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl,N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and/orheteroarylalkyl. “Substituted” further means any of the above groups inwhich one or more hydrogen atoms are replaced by a bond to an amino,cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkenyl,alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl,cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl,haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl,heteroaryl, N-heteroaryl and/or heteroarylalkyl group. In addition, eachof the foregoing substituents can also be optionally substituted withone or more of the above substituents.

As used herein, the symbol

(hereinafter can be referred to as “a point of attachment bond”) denotesa bond that is a point of attachment between two chemical entities, oneof which is depicted as being attached to the point of attachment bondand the other of which is not depicted as being attached to the point ofattachment bond. For example,

indicates that the chemical entity “XY” is bonded to another chemicalentity via the point of attachment bond. Furthermore, the specific pointof attachment to the non-depicted chemical entity can be specified byinference. For example, the compound CH₃—R³, wherein R³ is H or

infers that when R³ is “XY”, the point of attachment bond is the samebond as the bond by which R³ is depicted as being bonded to CH₃.

The following description includes information that may be useful inunderstanding the present invention. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed inventions, or that any publication specifically orimplicitly referenced is prior art.

Compounds of the Present Disclosure

The compound of the present disclosure can be useful for modulatingParkin ligase. Further, the compound of the present disclosure can beuseful for treating various diseases and conditions including, but notlimited to, cancer, neurological disease, a disorder characterized byabnormal accumulation of α-synuclein, a disorder of an aging process,cardiovascular disease, bacterial infection, viral infection,mitochondrial related disease, mental retardation, deafness, blindness,diabetes, obesity, autoimmune disease, glaucoma, Leber's HereditaryOptic Neuropathy, and rheumatoid arthritis. In one embodiment, thepresent disclosure provides compounds having the structure of formula(1):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L¹, L² and L³ are each independently selected from a bond, alkylene, oralkenylene;

M¹ and M² are each independently absent or independently selected from,—NR⁴—, —NR⁴C(O)—, —C(O)NR⁴—, —NR⁴C(O)NR⁴—, —C(O)—, —C(═NR⁴)—,—C(═NOR⁴)—, —OC(O)—, —C(O)O—, —OC(O)O—, —OC(O)NR⁴—, —NR⁴C(O)O—,—S(O)_(m)—, —S(O)_(m)NR⁴—, or —NR⁴S(O)_(m)—, provided that M¹ and M² arenot both —NR⁴—;

R¹, R², and R³ are each independently selected from an alkyl, alkenyl,cycloalkyl, aryl, biphenyl, heterocycloalkyl, heterocyclyl, heteroaryl,cycloalkylalkyl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclylalkyl,heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl, wherein eachcycloalkyl, aryl, heteroaryl, and heterocyclyl portion is optionallysubstituted with one or more R⁵;

R⁴ is each independently H, alkyl, wherein each alkyl is optionallysubstituted with one or more R⁵;

R⁵ is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH,NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶,SO₃H, SO₃R⁶, or SR⁶;

R⁶ is each independently alkyl or haloalkyl; or alternatively two R⁶ onthe same N atom can together form a 3-6 membered N-heterocyclyl; and

m is 0, 1, or 2.

In one embodiment, the compound of formula (1) is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and/or N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

Furthermore, in an embodiment wherein M¹ is absent, L¹ bonds directly toR¹ and in an embodiment wherein M² is absent, L² bonds directly to R².

In one embodiment, the present disclosure provides compounds having thestructure of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L¹, L² and L³ are each independently selected from a bond, alkylene, oralkenylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, —NR⁴C(O)NR⁴—, —C(O)—, —C(═NR⁴)—, —C(═NOR⁴)—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, —NR⁴C(O)O—, —S(O)_(m)—, —S(O)_(m)NR⁴—, or—NR⁴S(O)_(m)—, provided that M¹ and M² are not both —NR⁴—;

R¹, R², and R³ are each independently selected from an alkyl, alkenyl,cycloalkyl, aryl, biphenyl, heterocycloalkyl, heterocyclyl, heteroaryl,cycloalkylalkyl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclylalkyl,heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl, wherein eachcycloalkyl, aryl, heteroaryl, and heterocyclyl portion is optionallysubstituted with one or more R⁵;

R⁴ is each independently H, alkyl, wherein each alkyl is optionallysubstituted with one or more R⁵;

R⁵ is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH,NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶,SO₃H, SO₃R⁶, or SR⁶;

R⁶ is each independently alkyl or haloalkyl; or alternatively two R⁶ onthe same N atom can together form a 3-6 membered N-heterocyclyl; and

m is 0, 1, or 2.

In one embodiment, the compound of formula (I) is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and/or4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, L¹, L² and L³ in formula (I) are each independentlyselected from a bond, C1-C3 alkylene, or C2-C3 alkenylene. In someembodiments, L¹ is a bond. In another embodiment, L¹ is C1-C3 alkylene.In one embodiment, L¹ is C2-C3 alkenylene. In some embodiments, L² is abond. In another embodiment, L² is C1-C3 alkylene. In one embodiment, L²is C2-C3 alkenylene. In some embodiments, L³ is a bond. In anotherembodiment, L³ is C1-C3 alkylene. In one embodiment, L³ is C2-C3alkenylene. In one embodiment, L¹, L² and L³ in formula (I) are each abond.

In one embodiment, M¹ and M² in formula (I) are each independentlyselected from —NR⁴—, —NR⁴C(O)—, —C(O)NR⁴—, NR⁴C(O)NR⁴—, —OC(O)—,—C(O)O—, —OC(O)O—, —OC(O)NR⁴—, or —NR⁴C(O)O—. In another embodiment, M¹and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—. In one embodiment, R⁴ at each occurrence in the definition ofM¹ or M² is independently H or C1-C3 alkyl.

In one embodiment, M¹ and M² in formula (I) are each independentlyselected from —NH—, —N(CH₃)—, —NHC(O)—, —N(CH₃)C(O)—, —C(O)NH—, or—C(O)N(CH₃)—. In one embodiment, M¹ is —NH—. In another embodiment, M¹is —NHC(O)—. In some embodiments, M¹ is —N(CH₃)C(O)—. In one embodiment,M² is —NH—. In another embodiment, M² is —NHC(O)—. In some embodiments,M² is —N(CH₃)C(O)—.

In one embodiment, R¹, R², and R³ in formula (I) are each independentlyselected from alkyl, aryl, heteroaryl, heterocyclyl arylalkyl,arylalkenyl, heteroarylalkyl, heteroarylalkenyl, or heterocycloalkyl,wherein each cycloalkyl, aryl, heteroaryl and heterocyclyl portion isoptionally substituted with one or more R⁵. In another embodiment, R¹,R², and R³ are each independently selected from C1-C3 alkyl, phenyl,5-10 membered heteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl),5-6 membered heteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl),wherein each cycloalkyl, aryl, heteroaryl portion is optionallysubstituted with one or more R⁵.

In one embodiment, R¹ is an aryl, optionally substituted with one ormore R⁵. In another embodiment, R¹ is a phenyl, optionally substitutedwith one or more R⁵. In one embodiment, R¹ is an unsubstituted phenyl.In some embodiments, R¹ is a bicyclic aryl, optionally substituted withone or more R⁵. In one embodiment, R¹ is a

In some embodiments, R¹ is a heteroaryl, optionally substituted with oneor more R⁵. In some embodiments, R¹ is a pyridyl, optionally substitutedwith one or more R⁵. In some embodiments, R¹ is a furanyl, optionallysubstituted with one or more R⁵. In some embodiments, R¹ is athiophenyl, optionally substituted with one or more R⁵. In someembodiments, R¹ is a pyrimidinyl, optionally substituted with one ormore R⁵. In one embodiment, R¹ is a bicyclic heteroaryl, optionallysubstituted with one or more R⁵. In another embodiment, R¹ is

or optionally substituted with one or more R⁵. In another embodiment, R¹is

optionally substituted with one or more R⁵. In another embodiment, R¹ is

optionally substituted with one or more R⁵.

In one embodiment, R¹ is an arylalkyl, optionally substituted with oneor more R⁵. In some embodiments, R¹ is a phenylalkyl, optionallysubstituted with one or more R⁵. In another embodiment, R¹ is aphenyl-(C2-C3 alkyl), optionally substituted with one or more R⁵. In oneembodiment, R¹ is a phenyl-(C2 alkyl), optionally substituted with oneor more R⁵. In one embodiment, R¹ is an unsubstituted phenyl-(C2 alkyl).In one embodiment, R¹ is an arylalkenyl, optionally substituted with oneor more R⁵. In some embodiments, R¹ is a phenylalkenyl, optionallysubstituted with one or more R⁵. In another embodiment, R¹ is aphenyl-(C2-C3 alkenyl), optionally substituted with one or more R⁵. Inone embodiment, R¹ is a phenyl-(C2 alkenyl), optionally substituted withone or more R⁵. In one embodiment, R¹ is an unsubstituted phenyl-(C2alkenyl). In some embodiments, R¹ is a heterocyclyl, optionallysubstituted with one or more R⁵. In some embodiments, R¹ is apiperidinyl, optionally substituted with one or more R⁵. In someembodiments, R¹ is a tetrahydropyranyl, optionally substituted with oneor more R⁵.

In one embodiment, R² is an aryl, optionally substituted with one ormore R⁵. In another embodiment, R² is a phenyl, optionally substitutedwith one or more R⁵. In one embodiment, R² is an unsubstituted phenyl.In some embodiments, R² is a bicyclic aryl, optionally substituted withone or more R⁵. In one embodiments, R² is a

In some embodiments, R² is a heteroaryl, optionally substituted with oneor more R⁵. In some embodiments, R² is a pyridyl, optionally substitutedwith one or more R⁵. In some embodiments, R² is a furanyl, optionallysubstituted with one or more R⁵. In some embodiments, R² is athiophenyl, optionally substituted with one or more R⁵. In someembodiments, R² is a pyrimidinyl, optionally substituted with one ormore R⁵. In one embodiment, R² is a bicyclic heteroaryl, optionallysubstituted with one or more R⁵. In another embodiment, R² is

optionally substituted with one or more R⁵. In another embodiment, R² is

optionally substituted with one or more R⁵. In another embodiment, R² is

optionally substituted with one or more R⁵.

In one embodiment, R² is an arylalkyl, optionally substituted with oneor more R⁵. In some embodiments, R² is a phenylalkyl, optionallysubstituted with one or more R⁵. In another embodiment, R² is aphenyl-(C2-C3 alkyl), optionally substituted with one or more R⁵. In oneembodiment, R² is a phenyl-(C2 alkyl), optionally substituted with oneor more R⁵. In one embodiment, R² is an unsubstituted phenyl-(C2 alkyl).In one embodiment, R² is an arylalkenyl, optionally substituted with oneor more R⁵. In some embodiments, R² is a phenylalkenyl, optionallysubstituted with one or more R⁵. In another embodiment, R² is aphenyl-(C2-C3 alkenyl), optionally substituted with one or more R⁵. Inone embodiment, R² is a phenyl-(C2 alkenyl), optionally substituted withone or more R⁵. In one embodiment, R² is an unsubstituted phenyl-(C2alkenyl). In some embodiments, R² is a heterocyclyl, optionallysubstituted with one or more R⁵. In some embodiments, R² is apiperidinyl, optionally substituted with one or more R⁵. In someembodiments, R² is a tetrahydropyranyl, optionally substituted with oneor more R⁵.

In one embodiment, R³ is an aryl, optionally substituted with one ormore R⁵. In another embodiment, R³ is a phenyl, optionally substitutedwith one or more R⁵. In one embodiment, R³ is an unsubstituted phenyl.In some embodiments, R³ is a bicyclic aryl, optionally substituted withone or more R⁵. In one embodiments, R³ is a

In some embodiments, R³ is a heteroaryl, optionally substituted with oneor more R⁵. In some embodiments, R³ is a pyridyl, optionally substitutedwith one or more R⁵. In some embodiments, R³ is a furanyl, optionallysubstituted with one or more R⁵. In some embodiments, R³ is athiophenyl, optionally substituted with one or more R⁵. In someembodiments, R³ is a pyrimidinyl, optionally substituted with one ormore R⁵. In one embodiment, R³ is a bicyclic heteroaryl, optionallysubstituted with one or more R⁵. In another embodiment, R³ is

optionally substituted with one or more R⁵. In another embodiment, R³ is

optionally substituted with one or more R⁵. In another embodiment, R³ is

optionally substituted with one or more R⁵.

In one embodiment, R³ is an arylalkyl, optionally substituted with oneor more R⁵. In some embodiments, R³ is a phenylalkyl, optionallysubstituted with one or more R⁵. In another embodiment, R³ is aphenyl-(C2-C3 alkyl), optionally substituted with one or more R⁵. In oneembodiment, R³ is a phenyl-(C2 alkyl), optionally substituted with oneor more R⁵. In one embodiment, R³ is an unsubstituted phenyl-(C2 alkyl).In one embodiment, R³ is an arylalkenyl, optionally substituted with oneor more R⁵. In some embodiments, R³ is a phenylalkenyl, optionallysubstituted with one or more R⁵. In another embodiment, R³ is aphenyl-(C2-C3 alkenyl), optionally substituted with one or more R⁵. Inone embodiment, R³ is a phenyl-(C2 alkenyl), optionally substituted withone or more R⁵. In one embodiment, R³ is an unsubstituted phenyl-(C2alkenyl). In some embodiments, R³ is a heterocyclyl, optionallysubstituted with one or more R⁵. In some embodiments, R³ is apiperidinyl, optionally substituted with one or more R⁵. In someembodiments, R³ is a tetrahydropyranyl, optionally substituted with oneor more R⁵.

In one embodiment, R³ is alkyl. In another embodiment, R³ is methyl,ethyl, n-propyl, isopropyl, n-propyl, i-butyl, sec-butyl, or t-butyl. Inone embodiment, R³ is cycloalkyl. In another embodiment, R³ iscyclohexyl.

In some embodiments, at least one of R¹, R², and R³ in formula (I) isphenyl-(C2-C3 alkenyl), optionally substituted with one or more R⁵. Inanother embodiment, at least one of R¹, R², and R³ in formula (I) is aphenyl, optionally substituted with one or more R⁵. In otherembodiments, at least two of R¹, R², and R³ in formula (I) is a phenyl,optionally substituted with one or more R⁵.

In some embodiments, one or more of R¹, R², and R³ in formula (I) is

In one embodiment, R⁵ at each occurrence in the definition of R¹, R², orR³ is independently selected from I, Br, Cl, F, or C1-C6 alkyl. Inanother embodiment, R⁵ at each occurrence in the definition of R¹, R²,or R³ is independently selected from I, Br, Cl, F, or C1-C3 alkyl. Insome embodiments, R⁵ at each occurrence in the definition of R¹, R², orR³ is independently selected from I, Br, Cl, F, or methyl.

In one embodiment, R⁵ is each independently —CH₃, I, Br, Cl, F, CN, NH₂,NO₂, OH, OCF₃, OMe, —NMe₂, —NEt₂, or

In one embodiment, -L¹-M¹-R¹ or -L²-M²-R² of a compound of formula (I)are not —CH₂CH₂Ph.

In one embodiment, the compound of formula (I) has the structure offormula (I′):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³, M¹, M², R¹, R², and R³ are as defined above for formula (I).

In another embodiment, the compound of formula (I) has the structure offormula (I″):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is selected from a bond or C1-C3 alkylene;

M¹ and M² are each independently selected from —NR⁴—, —NR⁴C(O)— or—C(O)NR⁴—;

R¹, R², and R³ are each independently selected from phenyl, 5-10membered heteroaryl, phenyl-(C1-C3 alkyl), phenyl-(C2-C3 alkenyl), 5-6membered heteroaryl-(C1-C3 alkyl), or heteroaryl-(C2-C3 alkenyl),wherein each aryl or heteroaryl portion is optionally substituted withone or more R⁵;

R⁴ is each independently H or C1-C3 alkyl; and

R⁵ is each independently I, Br, Cl, F, or C1-C3 alkyl.

In one embodiment, the compound of formula (I), (I′), or (I″) is not

In one embodiment, the compound of formula (I) has the structure offormula (IA):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each phenyl, substituted with one or more R^(5a);

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5a) is each independently I, Br, Cl, F, C1-C6 alkyl, C1-C3 haloalkyl,—(C1-C6)-O—(C₁-C6), C₁-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (IB):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each phenyl, substituted with one or more R^(5a);

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5a) is each independently C1-C6 alkyl;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (IC):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each phenyl, substituted with one or more R⁵a, wherein atleast one of R¹ and R² is

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5a) is each independently I, Br, Cl, F, C1-C6 alkyl, C1-C3 haloalkyl,C1-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (ID):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each independently selected from —NR⁴C(O)— or —C(O)NR⁴—;

R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b);

R⁴ is each independently H or C1-C3 alkyl;

R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶,COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH,SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and

R⁶ is each independently alkyl or haloalkyl.

In one embodiment, the compound of formula (I) has the structure offormula (IE):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each —NHC(O)—;

R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b);

and R^(5b) is each independently I, Br, Cl, F, C1-C3 alkyl, C1-C3haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH.

In one embodiment, the compound of formula (I) has the structure offormula (IF):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each —NHC(O)—;

R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); and

R^(5b) is each independently C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy,C1-C3 haloalkoxy, OH, or COOH.

In one embodiment, the compound of formula (I) has the structure offormula (IG):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

L³ is a bond;

M¹ and M² are each —NHC(O)—;

R¹ and R² are each

R³ is phenyl; and

R^(5a) is each independently C1-C6 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy,C1-C3 haloalkoxy, OH, or COOH.

Various embodiments as described above for formula (I) also applies toformula (1), (I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), and (IG).

In one embodiment, the compound of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), or (IG) is notN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide,N-(3-benzamido-1-phenyl-1H-1,2,4-triazol-5-yl)furan-2-carboxamide,N-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,N-(1-phenyl-5-(phenylamino)-1H-1,2,4-triazol-3-yl)benzamide,4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and/or4-fluoro-N-(5-(4-methoxybenzamido)-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide,and N,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)bis(4-methylbenzamide).

In one embodiment, the compound of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), or (IG) is selected from Table 1 below, ora pharmaceutically acceptable salt or solvate thereof.

TABLE 1

A

G

J

B

C

D

E

H

K

F

M

I

L

U

V

W

X

Z

A1

B1

C1

D1

E1

F1

G1

H1

I1

J1

K1

L1

M1

N1

O1

U1

V1

W1

X1

Z1

A2

B2

C2

D2

E2

F2

G2

I2

J2

K2

L2

M2

O2

P2

R2

W2

X2

Y2

B3

D3

E3

F3

I3

J3

K3

L3

M3

N3

O3

P3

Q3

In one embodiment, the compound of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), or (IG) is selected from Table 2 below, ora pharmaceutically acceptable salt or solvate thereof.

TABLE 2

G

J

B

D

E

H

K

F

M

I

L

In one embodiment, the compound of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), and/or (IG) exclude

In one embodiment, the present disclosure provides compounds having thestructure of formula (2):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

M¹ and M² are each independently selected from a bond, —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—;

R¹ and R² are each independently selected from a cycloalkyl, aryl,heterocyclyl, or heteroaryl, wherein each cycloalkyl, aryl, heteroaryl,and heterocyclyl is optionally substituted with one or more R^(5a),provided that R¹ and R² are not 1,3-dioxoisoindolin-2-yl;

R³ is selected from an alkyl, alkenyl, cycloalkyl, aryl, heterocyclyl,or heteroaryl, wherein each cycloalkyl, aryl, heteroaryl, andheterocyclyl is optionally substituted with one or more R^(5a);

R⁴ is each independently H or alkyl;

R^(5a) is each independently I, Br, Cl, F, CN, NH₂, NHR^(6a), NO₂,NR^(6a)R^(6a), OH, OR^(6a), or R^(6a); and

R^(6a) is each independently alkyl or haloalkyl; or alternatively twoR^(6a) on the same N atom can together form a 3-6 memberedN-heterocyclyl.

In one embodiment, the present disclosure provides compounds having thestructure of formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

M¹ and M² are each independently selected from a bond, —NR⁴—, —NR⁴C(O)—,—C(O)NR⁴—, provided that M¹ and M² are not both —NR⁴— or both a bond;

R¹ and R² are each independently selected from a cycloalkyl, aryl,heterocyclyl, or heteroaryl, wherein each cycloalkyl, aryl, heteroaryl,and heterocyclyl is optionally substituted with one or more R^(5a),provided that R¹ and R² are not 1,3-dioxoisoindolin-2-yl;

R³ is selected from an alkyl, alkenyl, cycloalkyl, aryl, heterocyclyl,or heteroaryl, wherein each cycloalkyl, aryl, heteroaryl, andheterocyclyl is optionally substituted with one or more R^(5a);

R⁴ is each independently H or alkyl;

R^(5a) is each independently I, Br, Cl, F, CN, NH₂, NHR^(6a), NO₂,NR^(6a)R^(6a), OH, OR^(6a), or R^(6a); and

R^(6a) is each independently alkyl or haloalkyl; or alternatively twoR^(6a) on the same N atom can together form a 3-6 memberedN-heterocyclyl.

In one embodiment, M¹ in formula (II) is a bond. In another embodiment,M² in formula (II) is a bond. In one embodiment, M¹ in formula (II) is abond and M² is —NR⁴— or —NR⁴C(O)—. In one embodiment, M² in formula (II)is a bond and M¹ is —NR⁴— or —NR⁴C(O)—. In one embodiment, R⁴ at eachoccurrence in the definition of M¹ or M² is independently H or C1-C3alkyl.

In one embodiment, M¹ and M² in formula (I) are each independentlyselected from —NH—, —N(CH₃)—, —NHC(O)—, or —N(CH₃)C(O)—.

In one embodiment, R¹, R², and R³ in formula (I) are each independentlyselected from phenyl, 5-10 membered heteroaryl, or 5-10 memberedheterocyclyl, wherein each aryl, heteroaryl and heterocyclyl isoptionally substituted with one or more R^(5a).

In one embodiment, R¹ is an aryl, optionally substituted with one ormore R^(5a). In another embodiment, R¹ is a phenyl, optionallysubstituted with one or more R^(5a). In one embodiment, R¹ is anunsubstituted phenyl. In some embodiments, R¹ is a heteroaryl,optionally substituted with one or more R^(5a). In some embodiments, R¹is a pyridyl, optionally substituted with one or more R^(5a). In someembodiments, R¹ is a pyrimidinyl, optionally substituted with one ormore R^(5a). In one embodiment, R¹ is a bicyclic heteroaryl, optionallysubstituted with one or more R^(5a). In another embodiment, R¹ is

optionally substituted with one or more R^(5a).

In some embodiments, R¹ is a heterocyclyl, optionally substituted withone or more R^(5a). In some embodiments, R¹ is a tetrahydropyranyl,optionally substituted with one or more R^(5a).

In one embodiment, R² is an aryl, optionally substituted with one ormore R^(5a). In another embodiment, R² is a phenyl, optionallysubstituted with one or more R^(5a). In one embodiment, R² is anunsubstituted phenyl. In some embodiments, R² is a heteroaryl,optionally substituted with one or more R^(5a). In some embodiments, R²is a pyridyl, optionally substituted with one or more R^(5a). In someembodiments, R² is a pyrimidinyl, optionally substituted with one ormore R^(5a). In one embodiment, R² is a bicyclic heteroaryl, optionallysubstituted with one or more R^(5a). In another embodiment, R² is

optionally substituted with one or more R^(5a).

In some embodiments, R² is a heterocyclyl, optionally substituted withone or more R^(5a). In some embodiments, R² is a tetrahydropyranyl,optionally substituted with one or more R^(5a).

In one embodiment, R³ is an aryl, optionally substituted with one ormore R^(5a). In another embodiment, R³ is a phenyl, optionallysubstituted with one or more R^(5a). In one embodiment, R³ is anunsubstituted phenyl. In some embodiments, R³ is a pyridyl, optionallysubstituted with one or more R^(5a). In some embodiments, R³ is apyrimidinyl, optionally substituted with one or more R^(5a). In oneembodiment, R³ is a bicyclic heteroaryl, optionally substituted with oneor more R^(5a). In another embodiment, R³ is

optionally substituted with one or more R^(5a).

In some embodiments, at least one of R¹ or R² is a pyridyl, optionallysubstituted with one or more R^(5a). In other embodiments, at least oneof R¹ or R² is a 2-pyridyl, optionally substituted with one or moreR^(5a). In some embodiments, at least one of R¹ or R² is a pyridyl,substituted with at least one methyl and optionally with one or moreR^(5a). In one embodiment, R¹ is a pyridyl, optionally substituted withone or more R^(5a). In one embodiment, R² is a pyridyl, optionallysubstituted with one or more R^(5a).

In some embodiments, at least one of R¹ or R² is

optionally substituted with one or more R^(5a). In one embodiment,R^(5a) is C1-C3 alkyl. In another embodiment, R^(5a) is methyl.

In some embodiments, R¹ is

optionally substituted with one or more R^(5a). In one embodiment,R^(5a) is C1-C3 alkyl. In another embodiment, R^(5a) is methyl. In someembodiments, R² is

optionally substituted with one or more R^(5a). In one embodiment,R^(5a) is C1-C3 alkyl. In another embodiment, R^(5a) is methyl.

In some embodiments, at least one of R¹ or R² is

In some embodiments, R¹ is

In some embodiments, R² is

In some embodiments, R³ is a heterocyclyl, optionally substituted withone or more R^(5a). In some embodiments, R³ is a tetrahydropyranyl,optionally substituted with one or more R^(5a).

In one embodiment, R³ is alkyl. In one embodiment, R³ is C1-C6 alkyl. Inanother embodiment, R³ is methyl, ethyl, n-propyl, isopropyl, n-propyl,i-butyl, sec-butyl, or t-butyl. In one embodiment, R³ is cycloalkyl. Inanother embodiment, R³ is cyclohexyl.

In one embodiment, R^(5a) at each occurrence in the definition of R¹,R², or R³ is independently selected from I, Br, Cl, F, or C1-C6 alkyl.In another embodiment, R^(5a) at each occurrence in the definition ofR¹, R², or R³ is independently selected from I, Br, Cl, F, or C1-C3alkyl. In some embodiments, R^(5a) at each occurrence in the definitionof R¹, R², or R³ is independently selected from I, Br, Cl, F, or methyl.

In one embodiment, R^(5a) is each independently —CH₃, I, Br, Cl, F, CN,NH₂, NO₂, OH, OCF₃, OMe, —NMe₂, —NEt₂, or

In one embodiment, the compound of formula (1), (2), or (II) is selectedfrom Table 3 below, or a pharmaceutically acceptable salt or solvatethereof.

TABLE 3

T

Y1

Q2

S2

T2

U2

V2

A3

G3

H3

R3

S3

In another embodiment, the compounds described above may have particularfunctional characteristics. In one embodiment, the compound may have anoral bioavailability of about 10% to about 70% in a patient. In anotherembodiment, the compound may have an oral bioavailability of about 10%to about 50%. In another embodiment, the compound may have an oralbioavailability of about 10% to about 30%. In another embodiment, thecompound may have an oral bioavailability greater than about 20%. Inanother embodiment, the compound may have an oral bioavailability in apatient with any of the ranges above when administered in the assay asin Example 6.

In another embodiment, when administered orally, the compound may have aTmax of about 0.2 hrs to about 2 hrs in a patient. In anotherembodiment, the compound may have a Tmax of about 0.3 hrs to about 1 hrin a patient. In another embodiment, the compound may have a Tmax ofabout 0.4 hrs to about 0.6 hr in a patient. In another embodiment, thecompound may have a Tmax in a patient with any of the ranges above whenadministered in the assay as in Example 6.

In another embodiment, when administered orally, the compound may have aCmax of about 100 ng/mL to about 1,000 ng/mL in a patient. In anotherembodiment, when administered orally, the compound may have a Cmax ofabout 150 ng/mL to about 500 ng/mL in a patient. In another embodiment,when administered orally, the compound may have a Cmax of about 200ng/mL to about 400 ng/mL in a patient. In another embodiment, thecompound may have a Cmax in a patient with any of the ranges above whenadministered in the assay as in Example 6.

In another embodiment, the compound may have a half-life in human livermicrosomes greater than about 100 minutes. In another embodiment, thecompound may have a half-life in human liver microsomes greater thanabout 300 minutes. In another embodiment, the compound may have ahalf-life in human liver microsomes greater than about 500 minutes.

In another embodiment, the compound may have half-life in human livermicrosomes of about 100 minutes to about 1,000 minutes. In anotherembodiment, the compound may have half-life in human liver microsomes ofabout 200 minutes to about 800 minutes. In another embodiment, thecompound may have half-life in human liver microsomes of about 500minutes to about 700 minutes.

In another embodiment, the compound may have a half-life in rat livermicrosomes greater than about 100 minutes. In another embodiment, thecompound may have a half-life in rat liver microsomes greater than about300 minutes. In another embodiment, the compound may have a half-life inrat liver microsomes greater than about 500 minutes.

In another embodiment, the compound may have half-life in rat livermicrosomes of about 100 minutes to about 1,000 minutes. In anotherembodiment, the compound may have half-life in rat liver microsomes ofabout 200 minutes to about 800 minutes. In another embodiment, thecompound may have half-life in rat liver microsomes of about 500 minutesto about 700 minutes.

In a specific embodiment, the compound with any of the functionalcharacteristics as described above may be a compound of formula (1) (I),(I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II). In aspecific embodiment, the compound with the functional characteristics asdescribed above may from Table 1, Table 2 and/or Table 3.

Methods

Ubiquitination is crucial for a plethora of physiological processes,including cell survival and differentiation and innate and adaptiveimmunity. Proteins are built-up to cater for the structural andbiochemical requirements of the cell and they are also broken-down in ahighly-regulated process serving more purposes than just destruction andspace management. Proteins have different half-lives, determined by thenature of the amino acids present at their N-termini. Some will belong-lived, while other will rapidly be degraded. Proteolysis not onlyenables the cell to dispose of misfolded or damaged proteins, but alsoto fine-tune the concentration of essential proteins within the cell,such as the proteins involved in the cell cycle. This rapid, highlyspecific degradation can be achieved through the addition of one toseveral ubiquitin molecules to a target protein. The process is calledubiquitination.

In recent years, considerable progress has been made in theunderstanding of the molecular action of ubiquitin in signaling pathwaysand how alterations in the ubiquitin system lead to the development ofdistinct human diseases. It has been shown that ubiquitination plays arole in the onset and progression of cancer, metabolic syndromes,neurodegenerative diseases, autoimmunity, inflammatory disorders,infection and muscle dystrophies (Popovic et al. Nature Medicine 20,1242-1253 (2014)).

Ubiquitin-protein (E3) ligases are a large family of enzymes that selectvarious proteins for ubiquitination. These ubiquitin ligases, called “Ubligases” are known to have a role in various diseases and conditions,including but not limited to, cancer, inflammation and infectiousdiseases.

One specific Ub ligase is Parkin ligase. Parkin ligase is a component ofa multiprotein “E3” ubiquitin ligase complex, which in turn is part ofthe ubiquitin-proteasome system that mediates the targeting of proteinsfor degradation. Although the specific function of Parkin ligase is notknown, mutations in Parkin ligase are linked to various diseases, suchas Parkinson's disease, cancer and mycobacterial infection. Parkinligase is thus an attractive target for therapeutic intervention.

Further, there are various known methods for regulating ligases known inthe art. Many ligases, particularly ligases involved in theUbiquitin-Proteasome Pathway System (UPS), are known to have Zinc Finger(ZnF) domains that stabilize critical protein binding regions in thatligase.

ZnF domains coordinate zinc ions and this coordination stabilizesfunctional activity of the protein. The functional activity provided byproteins with ZnF domains can include the regulation of importantcellular signaling pathways, such as recognizing ubiquitins, regulationof DNA, such as transcription and repair, and acting as cellular redoxsensors. The binding of zinc to ZnF domains, or simply just regulatinghow zinc interacts with the ZnF domains, are essential to ligasesinvolved in the UPS.

Parkin ligase is known to have one or more ZnF domains. The presentdisclosure focuses on two different strategies for modulating ZnFdomains in Parkin ligase. One strategy of the present disclosureincludes using chelating compounds that bind to the ZnF domains and thusdisallowing the binding of zinc, or causing the dissociation of zinc,such as Zn, or Zn²⁺, from the ZnF domain. Another strategy of thepresent disclosure includes using compounds that bind or react with acysteine amino acid residue in the ZnF domain. One or more cysteineresidues (and sometimes with the assistance of histidine residues) areessential in ZnF domains for binding to and/or coordinating to the zincion. The zinc ion (usually Zn²⁺) can coordinate with multiple cysteineor histidine residues. The more cysteine residues there are in thedomain, the more flexible is the ZnF domain. Ligases, such as Parkinligase are thought to have multiple cysteine residues coordinated withzinc in their ZnF domains. This flexibility in the ZnF domains of Parkinligase is thought to allow the domain to be reversible, and is thus isone possible mechanism for regulating Parkin ligase. For example,efforts directed to this approach are disclosed in U.S. patentapplication Ser. No. 14/961,285; U.S. Provisional Application No.62/237,400; U.S. Provisional Application No. 62/222,008, and U.S.Provisional Application No. 62/087,972, all of which are herebyincorporated by reference in their entirety.

The present disclosure relates to the use of one or more agents or oneor more compounds of formula (1) (I), (I′), (I″), (IA), (IB), (IC),(ID), (IE), (IF), (IG), (2), or (II), or a pharmaceutically acceptablesalt or solvate thereof, which have electrophilic, chelation or bothelectrophilic and chelation properties that can interact with the zincion and/or the cysteine residue(s) in a Parkin ligase. In oneembodiment, compounds of the present disclosure modulate Parkin ligase'sactivity. Specifically, without bound to any theory, it is believed thatnot allowing a zinc ion to coordinate in at least one of Parkin ligase'sZnF domains induces its activity. The present disclosure is thusdirected to a method for activating or modulating Parkin ligase by thechelation of Zn followed by its removal from the ZnF domain, or throughelectrophilic attack at the cysteine amino acid(s) that holds the Zn inplace.

Accordingly, in one embodiment of the present disclosure, a method ofmodulating or activating a Parkin ligase comprising administering to asubject in need thereof a therapeutically effective amount of one ormore compounds of formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID),(IE), (IF), (IG), (2), or (II), or a pharmaceutically acceptable salt orsolvate thereof, is disclosed. In another embodiment, a method ofmodulating or activating a Parkin ligase comprising administering to asubject in need thereof a therapeutically effective amount of one ormore compounds of formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID),(IE), (IF), (IG), (2), or (II), or a pharmaceutically acceptable salt orsolvate thereof, that disrupt at least one Parkin ligase zinc finger isdisclosed. In another embodiment, a method of activating a Parkin ligasecomprising administering to a subject two or more compounds that disruptat least one Parkin ligase zinc finger is disclosed, wherein at leastone of the compound is selected from a compound of formula (1) (I),(I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or apharmaceutically acceptable salt or solvate thereof.

In a specific embodiment, the compounds of the present disclosure can bean electrophile or a chelator. In another embodiment, the compounds ofthe present disclosure can function as both an electrophile and as achelator. For example, the compounds of the present disclosure caninclude multiple functional groups wherein at least one functional grouphas chelating properties and at least one other functional group haselectrophilic properties.

In another specific embodiment, the compound useful for methods inmodulating or activating Parkin ligase as disclosed herein is selectedfrom Tables 1-3, or a pharmaceutically acceptable salt or solvatethereof.

In another embodiment, the compound of the present disclosure is usefulin a method to increase the Parkin ligase reaction with theActivity-based Ubiquitin vinyl sulfone probe. See e.g., Example 2.

In another embodiment, the one or more compounds of the presentdisclosure can coordinate with a Zn ion, and/or bind or react with oneor more cysteine residues. In a specific embodiment the Zn ion may beeither a Zn⁺ or a Zn²⁺ ion. In another embodiment, the compound cancoordinate to a Zn ion is a monodentate, bidentate, or tridentateligand.

In another embodiment, the compound of the present disclosure can bindand/or react with a thiol group in more than one cysteine residues. Inanother embodiment, the compound can bind and/or react with a thiolgroup in two cysteine residues. In another embodiment, the compound canbind and/or react with a thiol group in three cysteine residues. Inanother embodiment, the compound can bind and/or react with a thiolgroup in four cysteine residues. In another specific embodiment, thecompound can bind or react with one or more cysteine residues in one ormore domains selected from the group consisting amino acids 141-225,amino acids 238-293, amino acids 313-377, and amino acids 418-449 ofhuman Parkin ligase. See http://www.uniprot.org/uniprot/O60260.

The methods of the present disclosure also include activatingauto-ubiquitinization of a Parkin ligase by administering to a subjectin need thereof a therapeutically effective amount of one or morecompounds of formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE),(IF), (IG), (2), or (II), or a pharmaceutically acceptable salt orsolvate thereof.

In a specific embodiment, the one or more compounds of the presentdisclosure can disrupt at least one Parkin ligase zinc finger. Forexample, Phospho Ubiquitin (pUB), an endogenous cellular regulator ofParkin, can be added to Parkin ligase which can activate Parkin ligaseand its auto-ubiquitinization. In one embodiment, one or more compoundscan be administered to a subject in need thereof that actssynergistically with Phospho Ubiquitin (pUB) in activating the Parkinligase. See, e.g., Example 3. In one embodiment, the one or morecompounds that acts synergistically with pUB in activating the Parkinligase is a compound of formula (1) (I), (I′), (I″), (IA), (IB), (IC),(ID), (IE), (IF), (IG), (2), or (II), or a pharmaceutically acceptablesalt or solvate thereof. In another embodiment, one or more compounds ofthe present disclosure can be administered with pUB to synergisticallyincrease the activation of Parkin ligase and/or itsauto-ubiquitinization.

In another specific embodiment, the activation of the Parkin ligasetreats or reduces the incidence of one or more diseases or ailmentsselected from the group consisting of Alzheimer's Dementia, Parkinson'sdisease, Huntington Disease, Amyotrophic Lateral Sclerosis (ALS),Freidreich's ataxia, Spinocerebellar Ataxia, Multiple Systems Atrophy,PSP, Tauopathy, Diffuse Lewy Body Disease, Lewy Body dementia, anydisorder characterized by abnormal accumulation of α-synuclein,disorders of the aging process, stroke, bacterial infection, viralinfection, Mitochondrial related disease, mental retardation, deafness,blindness, diabetes, obesity, cardiovascular disease, multiplesclerosis, Sjogrens syndrome, lupus, glaucoma, includingpseudoexfoliation glaucoma, Leber's Hereditary Optic Neuropathy, andrheumatoid arthritis.

In a specific embodiment, the bacterial infection is Mycobacteriuminfection. In another specific embodiment the viral infection is HIV,Hepatitis B infection or Hepatitis C infection. Another embodiment ofthe present invention includes methods of treating and/or reducing theincidence of cancer, specifically comprising administering to a subjectin need thereof a therapeutically effective amount of one or morecompounds that disrupt at least one Parkin ligase zinc finger andinduces Parkin ligase activity. In a specific embodiment, the activatedParkin ligase suppresses the growth of one or more tumors and/orprevents metastasis of one or more tumors.

In another embodiment the cancer may be selected from one or more of thegroup consisting of Acute Lymphoblastic Leukemia, Acute MyeloidLeukemia, Adrenocortical Carcinoma, AIDS-Related Cancers, KaposiSarcoma, Lymphoma, Anal Cancer, Appendix Cancer, Astrocytomas, ChildhoodAtypical Teratoid/Rhabdoid Tumor, Basal Cell Carcinoma, Skin Cancer(Nonmelanoma), Childhood Bile Duct Cancer, Extrahepatic Bladder Cancer,Bone Cancer, Ewing Sarcoma Family of Tumors, Osteosarcoma and MalignantFibrous Histiocytoma, Brain Stem Glioma, Brain Tumors, Embryonal Tumors,Germ Cell Tumors, Craniopharyngioma, Ependymoma, Bronchial Tumors,Burkitt Lymphoma (Non-Hodgkin Lymphoma), Carcinoid Tumor,Gastrointestinal Carcinoma of Unknown Primary, Cardiac (Heart) Tumors,Lymphoma, Primary, Cervical Cancer, Childhood Cancers, Chordoma, ChronicLymphocytic Leukemia, Chronic Myelogenous Leukemia, ChronicMyeloproliferative Neoplasms Colon Cancer, Colorectal Cancer, CutaneousT-Cell Lymphoma, Ductal Carcinoma In Situ, Endometrial Cancer,Ependymoma, Esophageal Cancer, Esthesioneuroblastoma, Ewing Sarcoma,Extracranial Germ Cell Tumor, Extragonadal Germ Cell Tumor, ExtrahepaticBile Duct Cancer, Eye Cancer, Intraocular Melanoma, Retinoblastoma,Fibrous Histiocytoma of Bone, Malignant, and Osteosarcoma, GallbladderCancer, Gastric (Stomach) Cancer, Gastrointestinal Carcinoid Tumor,Gastrointestinal Stromal Tumors, Extragonadal Cancer, Ovarian Cancer,Testicular Cancer, Gestational Trophoblastic Disease, Glioma, Brain StemCancer, Hairy Cell Leukemia, Head and Neck Cancer, Heart Cancer,Hepatocellular (Liver) Cancer, Histiocytosis, Langerhans Cell Cancer,Hodgkin Lymphoma, Hypopharyngeal Cancer, Intraocular Melanoma, IsletCell Tumors, Pancreatic Neuroendocrine Tumors, Kaposi Sarcoma, KidneyCancer, Renal Cell Cancer, Wilms Tumor and Other Childhood KidneyTumors, Langerhans Cell Histiocytosis, Laryngeal Cancer, Leukemia,Chronic Lymphocytic Cancer, Chronic Myelogenous Cancer, Hairy CellCancer, Lip and Oral Cavity Cancer, Liver Cancer (Primary), LobularCarcinoma In Situ (LCIS), Lung Cancer, Non-Small Cell Cancer, Small CellCancer, Lymphoma, Cutaneous T-Cell (Mycosis Fungoides and SezarySyndrome), Hodgkin Cancer, Non-Hodgkin Cancer, Macroglobulinemia,Waldenström, Male Breast Cancer, Malignant Fibrous Histiocytoma of Boneand Osteosarcoma, Melanoma, Intraocular (Eye) Cancer, Merkel CellCarcinoma, Mesothelioma, Malignant, Metastatic Squamous Neck Cancer withOccult Primary, Midline Tract Carcinoma Involving NUT Gene, MouthCancer, Multiple Endocrine Neoplasia Syndromes, Multiple Myeloma/PlasmaCell Neoplasm, Mycosis Fungoides, Myelodysplastic Syndromes,Myelodysplastic/Myeloproliferative Neoplasms, Myelogenous Leukemia,Chronic, Myeloid Leukemia, Acute, Myeloma Multiple, ChronicMyeloproliferative Neoplasms, Nasal Cavity and Paranasal Sinus Cancer,Nasopharyngeal Cancer, Neuroblastoma, Non-Hodgkin Lymphoma, Non-SmallCell Lung Cancer, Oral Cancer, Oral Cavity Cancer, Lip and OropharyngealCancer, Osteosarcoma and Malignant Fibrous Histiocytoma of Bone,Epithelial Cancer, Low Malignant Potential Tumor, Pancreatic Cancer,Pancreatic Neuroendocrine Tumors (Islet Cell Tumors), Papillomatosis,Paraganglioma, Parathyroid Cancer, Penile Cancer, Pharyngeal Cancer,Pheochromocytoma, Pituitary Tumor, Plasma Cell Neoplasm/MultipleMyeloma, Pleuropulmonary Blastoma, Primary Central Nervous SystemLymphoma, Rectal Cancer, Renal Cell (Kidney) Cancer, Retinoblastoma,Rhabdomyosarcoma, Salivary Gland Cancer, Sarcoma, Ewing Cancer, KaposiCancer, Osteosarcoma (Bone Cancer), Soft Tissue Cancer, Uterine Cancer,Sezary Syndrome, Skin Cancer, Childhood Melanoma, Merkel Cell Carcinoma,Nonmelanoma, Small Cell Lung Cancer, Small Intestine Cancer, Soft TissueSarcoma, Squamous Cell Carcinoma, Skin Cancer (Nonmelanoma), ChildhoodSquamous Neck Cancer with Occult Primary, Metastatic Cancer, Stomach(Gastric) Cancer, T-Cell Lymphoma, Cutaneous Cancer, Testicular Cancer,Throat Cancer, Thymoma and Thymic Carcinoma, Thyroid Cancer,Transitional Cell Cancer of the Renal Pelvis and Ureter, UnknownPrimary, Carcinoma of Childhood, Unusual Cancers of Childhood, UrethralCancer, Uterine Cancer, Endometrial Cancer, Uterine Sarcoma, VaginalCancer, Vulvar Cancer, Waldenström Macroglobulinemia, Wilms Tumor, andWomen's Cancers.

In a specific embodiment, the cancer is glioblastoma, small cell lungcarcinoma, breast cancer and/or prostate cancer. In another embodiment,the administration of the Parkin ligase suppresses one or more tumors inthe subject.

In another specific embodiment, the compound eliminates damagedmitochondria, increases cell viability during cellular stress, decreasestumor transformation and/or mitigates alpha-synuclein in cells.

In another embodiment, the methods of the present disclosure includetreating and/or reducing the incidence of Parkinson's disease,specifically by administering to a subject in need thereof atherapeutically effective amount of one or more compounds that disruptat least one Parkin ligase zinc finger and induces Parkin ligaseactivity, wherein the compound can coordinate with a Zn ion and/or reactwith a thiol group in a cysteine(s). In one embodiment, the compoundthat disrupts at least one Parkin ligase zinc finger and includes Parkinligase activity in the above mentioned method is selected from compoundof formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF),(IG), (2), or (II), or a pharmaceutically acceptable salt or solvatethereof. In another embodiment, the one or more compounds eliminatedamaged mitochondria, increases cell viability during cellular stressand/or mitigates alpha-synuclein in cells. “Somatic Mutations of theParkinson's disease-associated gene PARK2 in glioblastoma and otherhuman malignancies” (Nature Genetics January 2010 42(1)77-82). In oneembodiment, the compound that eliminate damaged mitochondria, increasecell viability during cellular stress and/or mitigates alpha-synucleinin cells in the above mentioned method is a selected from compound offormula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG),(2), or (II), or a pharmaceutically acceptable salt or solvate thereof.

In another embodiment, the Parkin ligase activation altersubiquitination. Specifically, the alteration of ubiquitination is causedby the ability of Parkin to modify a substrate protein by covalentattachment of Ubiquitin, a substrate protein being Parkin itself, oranother protein such as Mitofusion 1 or 2, FBW7, or other publiclyreported substrates of Parkin ligase.

Further embodiments of the present disclosure relate to methods oftreating, preventing, or ameliorating one or more symptoms associatedwith neurological diseases or disorders including but not limited toAlzheimer's Dementia, Parkinson's disease, Huntington Disease,Amyotrophic Lateral Sclerosis (ALS), Freidreich's ataxia,Spinocerebellar Ataxia, Multiple Systems Atrophy, PSP, Tauopathy,Diffuse Lewy Body Disease, Lewy Body dementia, any disordercharacterized by abnormal accumulation of α-synuclein, disorders of theaging process, and stroke.

Other embodiments of the present disclosure relate to methods oftreating, preventing, or ameliorating one or more symptoms associatedwith but not limited to mental retardation, deafness, blindness,diabetes, obesity, cardiovascular disease, and autoimmune diseases suchas multiple sclerosis, Sjogrens syndrome, lupus, glaucoma, includingpseudoexfoliation glaucoma, Leber's Hereditary Optic Neuropathy, andrheumatoid arthritis.

Further embodiments of the present disclosure of the present inventionrelate to methods of treating, preventing, or ameliorating one or moresymptoms associated with but not limited to Mitochondrial RelatedDiseases or Capsules as follows:

-   -   Alpers Disease    -   Barth Syndrome/LIC (Lethal Infantile Cardiomyopathy)    -   Beta-oxidation Defects    -   Carnitine-Acyl-Carnitine Deficiency    -   Carnitine Deficiency    -   Creatine Deficiency Syndromes    -   Co-Enzyme Q10 Deficiency    -   Complex I Deficiency    -   Complex II Deficiency    -   Complex III Deficiency    -   Complex IV Deficiency/COX Deficiency    -   Complex V Deficiency    -   CPEO    -   CPT I Deficiency    -   CPT II Deficiency    -   KSS    -   Lactic Acidosis    -   LBSL—Leukodystrohpy    -   LCAD    -   LCHAD    -   Leigh Disease or Syndrome    -   Luft Disease    -   MAD/Glutaric Aciduria Type II    -   MCAD    -   MELAS    -   MERRF    -   MIRAS    -   Mitochondrial Cytopathy    -   Mitochondrial DNA Depletion    -   Mitochondrial Encephalopathy    -   Mitochondrial Myopathy    -   MNGIE    -   NARP    -   Pearson Syndrome    -   Pyruvate Carboxylase Deficiency    -   Pyruvate Dehydrogenase Deficiency    -   POLG Mutations    -   Respiratory Chain    -   SCAD    -   SCHAD    -   VLCAD.

In one embodiment, the methods of the present disclosure includetreating and/or reducing the incidence of cancer, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of formula (1) (I), (I′), (I″), (IA), (IB), (IC),(ID), (IE), (IF), (IG), (2), or (II), or a pharmaceutically acceptablesalt or solvate thereof. The compound of the present disclosure candisrupts at least one Parkin ligase zinc finger and induces Parkinligase activity, wherein the compound can coordinate with a zinc ionand/or bind or react with a cysteine. In a specific embodiment, theParkin ligase suppresses the growth of one or more tumors and/orprevents metastasis of one or more tumors. In another embodiment, thecompound of formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE),(IF), (IG), (2), or (II), or a pharmaceutically acceptable salt orsolvate thereof eliminates damaged mitochondria, increases cellviability during cellular stress, decreases tumor transformation and/ormitigates alpha-synuclein in cells. In another embodiment, the cancer isglioblastoma, small cell lung carcinoma, breast cancer or prostatecancer.

In a specific embodiment, the methods of the present disclosure includetreating and/or reducing the incidence of Parkinson's disease,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or a pharmaceuticallyacceptable salt or solvate thereof that disrupts at least one Parkinligase zinc finger and induces Parkin ligase activity, wherein thecompound can coordinate with a zinc ion and/or bind or react with acysteine. In a specific embodiment, the compound of the presentdisclosure eliminates damaged mitochondria, increases cell viabilityduring cellular stress and/or mitigates alpha-synuclein in cells.

Pharmaceutical Compositions and Formulations

The present disclosure also includes pharmaceutical compositions formodulating or activating a Parkin ligase in a subject. In oneembodiment, a pharmaceutical composition comprises one or more compoundsof formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF),(IG), (2), or (II), or a pharmaceutically acceptable salt or solvatethereof. In some embodiments, one or more compounds of formula (1) (I),(I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or apharmaceutically acceptable salt or solvate thereof, in a pharmaceuticalcomposition as described herein disrupts at least one Parkin ligase zincfinger. In another embodiment, one or more compounds of formula (1) (I),(I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or apharmaceutically acceptable salt or solvate thereof, in a pharmaceuticalcomposition as described herein coordinates with a Zn ion, and/or reactwith at least one thiol group in a cysteine.

In one embodiment of the present disclosure, a pharmaceuticalcomposition comprises a therapeutically effective amounts of one or morecompounds of formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE),(IF), (IG), (2), or (II), or a pharmaceutically acceptable salt orsolvate thereof.

In a specific embodiment, a pharmaceutical composition, as describedherein, comprises one or more compounds selected from Table 1, or apharmaceutically acceptable salt or solvate thereof. In one embodiment,a pharmaceutical composition as described herein comprise one or morecompounds selected from Table 2, or a pharmaceutically acceptable saltor solvate thereof. In one embodiment, a pharmaceutical composition asdescribed herein comprise one or more compounds selected from Table 3,or a pharmaceutically acceptable salt or solvate thereof.

In one embodiment, a pharmaceutical composition described herein doesnot contain:

In one embodiment, a pharmaceutical composition, as described herein,comprising one or more compounds of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or a pharmaceuticallyacceptable salt or solvate thereof, further comprises one or moreadditional therapeutically active agents. In one embodiment, one or moreadditional therapeutically active agents are selected from therapeuticsuseful for treating cancer, neurological disease, a disordercharacterized by abnormal accumulation of α-synuclein, a disorder of anaging process, cardiovascular disease, bacterial infection, viralinfection, mitochondrial related disease, mental retardation, deafness,blindness, diabetes, obesity, autoimmune disease, glaucoma, Leber'sHereditary Optic Neuropathy, and rheumatoid arthritis.

In a further embodiment of the present disclosure, a pharmaceuticalcomposition comprising one or more compounds of formula (1) (I), (I′),(I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or apharmaceutically acceptable salt or solvate thereof, and apharmaceutically acceptable excipient or adjuvant is provided. Thepharmaceutically acceptable excipients and adjuvants are added to thecomposition or formulation for a variety of purposes. In anotherembodiment, a pharmaceutical composition comprising one or morecompounds of formula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE),(IF), (IG), (2), or (II), or a pharmaceutically acceptable salt orsolvate thereof, further comprises a pharmaceutically acceptablecarrier. In one embodiment, a pharmaceutically acceptable carrierincludes a pharmaceutically acceptable excipient, binder, and/ordiluent. In one embodiment, suitable pharmaceutically acceptableexcipients include, but are not limited to, water, salt solutions,alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesiumstearate, talc, silicic acid, viscous paraffin, hydroxymethylcelluloseand polyvinylpyrrolidone.

In certain embodiments, the pharmaceutical compositions of the presentdisclosure may additionally contain other adjunct componentsconventionally found in pharmaceutical compositions, at theirart-established usage levels. Thus, for example, the pharmaceuticalcompositions may contain additional, compatible, pharmaceutically-activematerials such as, for example, antipruritics, astringents, localanesthetics or anti-inflammatory agents, or may contain additionalmaterials useful in physically formulating various dosage forms of thecompositions of the present invention, such as dyes, flavoring agents,preservatives, antioxidants, opacifiers, thickening agents andstabilizers. However, such materials, when added, should not undulyinterfere with the biological activities of the components of thecompositions of the present invention. The formulations can besterilized and, if desired, mixed with auxiliary agents, e.g.,lubricants, preservatives, stabilizers, wetting agents, emulsifiers,salts for influencing osmotic pressure, buffers, colorings, flavoringsand/or aromatic substances and the like which do not deleteriouslyinteract with the oligonucleotide(s) of the formulation.

For the purposes of this disclosure, the compounds of the presentdisclosure can be formulated for administration by a variety of meansincluding orally, parenterally, by inhalation spray, topically, orrectally in formulations containing pharmaceutically acceptablecarriers, adjuvants and vehicles. The term parenteral as used hereincludes subcutaneous, intravenous, intramuscular, and intraarterialinjections with a variety of infusion techniques. Intraarterial andintravenous injection as used herein includes administration throughcatheters.

The compounds disclosed herein can be formulated in accordance with theroutine procedures adapted for desired administration route.Accordingly, the compounds disclosed herein can take such forms assuspensions, solutions or emulsions in oily or aqueous vehicles, and cancontain formulatory agents such as suspending, stabilizing and/ordispersing agents. The compounds disclosed herein can also be formulatedas a preparation for implantation or injection.

Thus, for example, the compounds can be formulated with suitablepolymeric or hydrophobic materials (e.g., as an emulsion in anacceptable oil) or ion exchange resins, or as sparingly solublederivatives (e.g., as a sparingly soluble salt). Alternatively, theactive ingredient can be in powder form for constitution with a suitablevehicle, e.g., sterile pyrogen-free water, before use. Suitableformulations for each of these methods of administration can be found,for example, in Remington: The Science and Practice of Pharmacy, A.Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins,Philadelphia, Pa.

In certain embodiments, a pharmaceutical composition of the presentdisclosure is prepared using known techniques, including, but notlimited to mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or tableting processes.

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising a compound of formula (1) (I), (I′), (I″), (IA),(IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or a pharmaceuticallyacceptable salt or solvate thereof, as disclosed herein, combined with apharmaceutically acceptable carrier. In one embodiment, suitablepharmaceutically acceptable carriers include, but are not limited to,inert solid fillers or diluents and sterile aqueous or organicsolutions. Pharmaceutically acceptable carriers are well known to thoseskilled in the art and include, but are not limited to, from about 0.01to about 0.1 M and preferably 0.05M phosphate buffer or 0.8% saline.Such pharmaceutically acceptable carriers can be aqueous or non-aqueoussolutions, suspensions and emulsions. Examples of non-aqueous solventssuitable for use in the present application include, but are not limitedto, propylene glycol, polyethylene glycol, vegetable oils such as oliveoil, and injectable organic esters such as ethyl oleate.

Aqueous carriers suitable for use in the present application include,but are not limited to, water, ethanol, alcoholic/aqueous solutions,glycerol, emulsions or suspensions, including saline and buffered media.Oral carriers can be elixirs, syrups, capsules, tablets and the like.

Liquid carriers suitable for use in the present application can be usedin preparing solutions, suspensions, emulsions, syrups, elixirs andpressurized compounds. The active ingredient can be dissolved orsuspended in a pharmaceutically acceptable liquid carrier such as water,an organic solvent, a mixture of both or pharmaceutically acceptableoils or fats. The liquid carrier can contain other suitablepharmaceutical additives such as solubilizers, emulsifiers, buffers,preservatives, sweeteners, flavoring agents, suspending agents,thickening agents, colors, viscosity regulators, stabilizers orosmo-regulators.

Liquid carriers suitable for use in the present application include, butare not limited to, water (partially containing additives as above, e.g.cellulose derivatives, preferably sodium carboxymethyl cellulosesolution), alcohols (including monohydric alcohols and polyhydricalcohols, e.g. glycols) and their derivatives, and oils (e.g.fractionated coconut oil and arachis oil). For parenteraladministration, the carrier can also include an oily ester such as ethyloleate and isopropyl myristate. Sterile liquid carriers are useful insterile liquid form comprising compounds for parenteral administration.The liquid carrier for pressurized compounds disclosed herein can behalogenated hydrocarbon or other pharmaceutically acceptable propellent.

Solid carriers suitable for use in the present application include, butare not limited to, inert substances such as lactose, starch, glucose,methyl-cellulose, magnesium stearate, dicalcium phosphate, mannitol andthe like. A solid carrier can further include one or more substancesacting as flavoring agents, lubricants, solubilizers, suspending agents,fillers, glidants, compression aids, binders or tablet-disintegratingagents; it can also be an encapsulating material. In powders, thecarrier can be a finely divided solid which is in admixture with thefinely divided active compound. In tablets, the active compound is mixedwith a carrier having the necessary compression properties in suitableproportions and compacted in the shape and size desired. The powders andtablets preferably contain up to 99% of the active compound. Suitablesolid carriers include, for example, calcium phosphate, magnesiumstearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose,polyvinylpyrrolidine, low melting waxes and ion exchange resins. Atablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active ingredient in a freeflowing form such as a powder or granules, optionally mixed with abinder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (e.g., sodiumstarch glycolate, cross-linked povidone, cross-linked sodiumcarboxymethyl cellulose) surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsmay optionally be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropyl methylcellulose in varying proportionsto provide the desired release profile. Tablets may optionally beprovided with an enteric coating, to provide release in parts of the gutother than the stomach.

Parenteral carriers suitable for use in the present application include,but are not limited to, sodium chloride solution, Ringer's dextrose,dextrose and sodium chloride, lactated Ringer's and fixed oils.Intravenous carriers include fluid and nutrient replenishers,electrolyte replenishers such as those based on Ringer's dextrose andthe like. Preservatives and other additives can also be present, suchas, for example, antimicrobials, antioxidants, chelating agents, inertgases and the like.

Carriers suitable for use in the present application can be mixed asneeded with disintegrants, diluents, granulating agents, lubricants,binders and the like using conventional techniques known in the art. Thecarriers can also be sterilized using methods that do not deleteriouslyreact with the compounds, as is generally known in the art.

Diluents may be added to the formulations of the present invention.Diluents increase the bulk of a solid pharmaceutical composition and/orcombination, and may make a pharmaceutical dosage form containing thecomposition and/or combination easier for the patient and care giver tohandle. Diluents for solid compositions and/or combinations include, forexample, microcrystalline cellulose (e.g., AVICEL), microfine cellulose,lactose, starch, pregelatinized starch, calcium carbonate, calciumsulfate, sugar, dextrates, dextrin, dextrose, dibasic calcium phosphatedihydrate, tribasic calcium phosphate, kaolin, magnesium carbonate,magnesium oxide, maltodextrin, mannitol, polymethacrylates (e.g.,EUDRAGIT(r)), potassium chloride, powdered cellulose, sodium chloride,sorbitol, and talc.

Additional embodiments relate to the pharmaceutical formulations whereinthe formulation is selected from the group consisting of a solid,powder, liquid and a gel. In certain embodiments, a pharmaceuticalcomposition of the present invention is a solid (e.g., a powder, tablet,a capsule, granulates, and/or aggregates). In certain of suchembodiments, a solid pharmaceutical composition comprising one or moreingredients known in the art, including, but not limited to, starches,sugars, diluents, granulating agents, lubricants, binders, anddisintegrating agents.

Solid pharmaceutical compositions that are compacted into a dosage form,such as a tablet, may include excipients whose functions include helpingto bind the active ingredient and other excipients together aftercompression. Binders for solid pharmaceutical compositions and/orcombinations include acacia, alginic acid, carbomer (e.g., carbopol),carboxymethylcellulose sodium, dextrin, ethyl cellulose, gelatin, guargum, gum tragacanth, hydrogenated vegetable oil, hydroxyethyl cellulose,hydroxypropyl cellulose (e.g., KLUCEL), hydroxypropyl methyl cellulose(e.g., METHOCEL), liquid glucose, magnesium aluminum silicate,maltodextrin, methylcellulose, polymethacrylates, povidone (e.g.,KOLLIDON, PLASDONE), pregelatinized starch, sodium alginate, and starch.

The dissolution rate of a compacted solid pharmaceutical composition inthe patient's stomach may be increased by the addition of a disintegrantto the composition and/or combination. Disintegrants include alginicacid, carboxymethylcellulose calcium, carboxymethylcellulose sodium(e.g., AC-DI-SOL and PRIMELLOSE), colloidal silicon dioxide,croscarmellose sodium, crospovidone (e.g., KOLLIDON and POLYPLASDONE),guar gum, magnesium aluminum silicate, methyl cellulose,microcrystalline cellulose, polacrilin potassium, powdered cellulose,pregelatinized starch, sodium alginate, sodium starch glycolate (e.g.,EXPLOTAB), potato starch, and starch.

Glidants can be added to improve the flowability of a non-compactedsolid composition and/or combination and to improve the accuracy ofdosing. Excipients that may function as glidants include colloidalsilicon dioxide, magnesium trisilicate, powdered cellulose, starch,talc, and tribasic calcium phosphate.

When a dosage form such as a tablet is made by the compaction of apowdered composition, the composition is subjected to pressure from apunch and dye. Some excipients and active ingredients have a tendency toadhere to the surfaces of the punch and dye, which can cause the productto have pitting and other surface irregularities. A lubricant can beadded to the composition and/or combination to reduce adhesion and easethe release of the product from the dye. Lubricants include magnesiumstearate, calcium stearate, glyceryl monostearate, glycerylpalmitostearate, hydrogenated castor oil, hydrogenated vegetable oil,mineral oil, polyethylene glycol, sodium benzoate, sodium laurylsulfate, sodium stearyl fumarate, stearic acid, talc, and zinc stearate.

Flavoring agents and flavor enhancers make the dosage form morepalatable to the patient. Common flavoring agents and flavor enhancersfor pharmaceutical products that may be included in the compositionand/or combination of the present invention include maltol, vanillin,ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol, andtartaric acid.

Solid and liquid compositions may also be dyed using anypharmaceutically acceptable colorant to improve their appearance and/orfacilitate patient identification of the product and unit dosage level.

In certain embodiments, a pharmaceutical composition of the presentinvention is a liquid (e.g., a suspension, elixir and/or solution). Incertain of such embodiments, a liquid pharmaceutical composition isprepared using ingredients known in the art, including, but not limitedto, water, glycols, oils, alcohols, flavoring agents, preservatives, andcoloring agents.

Liquid pharmaceutical compositions can be prepared using compounds offormula (1) (I), (I′), (I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG),(2), or (II), or a pharmaceutically acceptable salt or solvate thereof,and any other solid excipients where the components are dissolved orsuspended in a liquid carrier such as water, vegetable oil, alcohol,polyethylene glycol, propylene glycol, or glycerin.

For example, formulations for parenteral administration can contain ascommon excipients sterile water or saline, polyalkylene glycols such aspolyethylene glycol, oils of vegetable origin, hydrogenated naphthalenesand the like. In particular, biocompatible, biodegradable lactidepolymer, lactide/glycolide copolymer, orpolyoxyethylene-polyoxypropylene copolymers can be useful excipients tocontrol the release of active compounds. Other potentially usefulparenteral delivery systems include ethylene-vinyl acetate copolymerparticles, osmotic pumps, implantable infusion systems, and liposomes.Formulations for inhalation administration contain as excipients, forexample, lactose, or can be aqueous solutions containing, for example,polyoxyethylene-9-auryl ether, glycocholate and deoxycholate, or oilysolutions for administration in the form of nasal drops, or as a gel tobe applied intranasally. Formulations for parenteral administration canalso include glycocholate for buccal administration, methoxysalicylatefor rectal administration, or citric acid for vaginal administration.

Liquid pharmaceutical compositions can contain emulsifying agents todisperse uniformly throughout the composition and/or combination anactive ingredient or other excipient that is not soluble in the liquidcarrier. Emulsifying agents that may be useful in liquid compositionsand/or combinations of the present invention include, for example,gelatin, egg yolk, casein, cholesterol, acacia, tragacanth, chondrus,pectin, methyl cellulose, carbomer, cetostearyl alcohol, and cetylalcohol.

Liquid pharmaceutical compositions can also contain a viscosityenhancing agent to improve the mouth-feel of the product and/or coat thelining of the gastrointestinal tract. Such agents include acacia,alginic acid bentonite, carbomer, carboxymethylcellulose calcium orsodium, cetostearyl alcohol, methyl cellulose, ethylcellulose, gelatinguar gum, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, maltodextrin, polyvinyl alcohol, povidone, propylenecarbonate, propylene glycol alginate, sodium alginate, sodium starchglycolate, starch tragacanth, and xanthan gum.

Sweetening agents such as aspartame, lactose, sorbitol, saccharin,sodium saccharin, sucrose, aspartame, fructose, mannitol, and invertsugar may be added to improve the taste.

Preservatives and chelating agents such as alcohol, sodium benzoate,butylated hydroxyl toluene, butylated hydroxyanisole, andethylenediamine tetraacetic acid may be added at levels safe foringestion to improve storage stability.

A liquid composition can also contain a buffer such as guconic acid,lactic acid, citric acid or acetic acid, sodium guconate, sodiumlactate, sodium citrate, or sodium acetate. Selection of excipients andthe amounts used may be readily determined by the formulation scientistbased upon experience and consideration of standard procedures andreference works in the field.

In one embodiment, a pharmaceutical composition is prepared foradministration by injection (e.g., intravenous, subcutaneous,intramuscular, etc.). In certain of such embodiments, a pharmaceuticalcomposition comprises a carrier and is formulated in aqueous solution,such as water or physiologically compatible buffers such as Hanks'ssolution, Ringer's solution, or physiological saline buffer. In certainembodiments, other ingredients are included (e.g., ingredients that aidin solubility or serve as preservatives). In certain embodiments,injectable suspensions are prepared using appropriate liquid carriers,suspending agents and the like. Certain pharmaceutical compositions forinjection are presented in unit dosage form, e.g., in ampoules or inmulti-dose containers. Certain pharmaceutical compositions for injectionare suspensions, solutions or emulsions in oily or aqueous vehicles, andmay contain formulatory agents such as suspending, stabilizing and/ordispersing agents. Certain solvents suitable for use in pharmaceuticalcompositions for injection include, but are not limited to, lipophilicsolvents and fatty oils, such as sesame oil, synthetic fatty acidesters, such as ethyl oleate or triglycerides, and liposomes. Aqueousinjection suspensions may contain substances that increase the viscosityof the suspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, such suspensions may also contain suitablestabilizers or agents that increase the solubility of the pharmaceuticalagents to allow for the preparation of highly concentrated solutions.

The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, such as a solution in 1,3-butane-diol or prepared as alyophilized powder. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile fixed oils may conventionally be employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid may likewise be used in the preparationof injectables. Formulations for intravenous administration can comprisesolutions in sterile isotonic aqueous buffer. Where necessary, theformulations can also include a solubilizing agent and a localanesthetic to ease pain at the site of the injection. Generally, theingredients are supplied either separately or mixed together in unitdosage form, for example, as a dry lyophilized powder or water freeconcentrate in a hermetically sealed container such as an ampule orsachet indicating the quantity of active agent. Where the compound is tobe administered by infusion, it can be dispensed in a formulation withan infusion bottle containing sterile pharmaceutical grade water, salineor dextrose/water. Where the compound is administered by injection, anampule of sterile water for injection or saline can be provided so thatthe ingredients can be mixed prior to administration.

Suitable formulations further include aqueous and non-aqueous sterileinjection solutions that can contain antioxidants, buffers,bacteriostats, bactericidal antibiotics and solutes that render theformulation isotonic with the bodily fluids of the intended recipient;and aqueous and non-aqueous sterile suspensions, which can includesuspending agents and thickening agents.

In certain embodiments, a pharmaceutical composition of the presentinvention is formulated as a depot preparation. Certain such depotpreparations are typically longer acting than non-depot preparations. Incertain embodiments, such preparations are administered by implantation(for example subcutaneously or intramuscularly) or by intramuscularinjection. In certain embodiments, depot preparations are prepared usingsuitable polymeric or hydrophobic materials (for example an emulsion inan acceptable oil) or ion exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises a delivery system. Examples of delivery systemsinclude, but are not limited to, liposomes and emulsions. Certaindelivery systems are useful for preparing certain pharmaceuticalcompositions including those comprising hydrophobic compounds. Incertain embodiments, certain organic solvents such as dimethylsulfoxideare used.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises a co-solvent system. Certain of such co-solventsystems comprise, for example, benzyl alcohol, a nonpolar surfactant, awater-miscible organic polymer, and an aqueous phase. In certainembodiments, such co-solvent systems are used for hydrophobic compounds.A non-limiting example of such a co-solvent system is the VPD co-solventsystem, which is a solution of absolute ethanol comprising 3% w/v benzylalcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/vpolyethylene glycol 300. The proportions of such co-solvent systems maybe varied considerably without significantly altering their solubilityand toxicity characteristics. Furthermore, the identity of co-solventcomponents may be varied: for example, other surfactants may be usedinstead of Polysorbate 80; the fraction size of polyethylene glycol maybe varied; other biocompatible polymers may replace polyethylene glycol,e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides maysubstitute for dextrose.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises a sustained-release system. A non-limiting exampleof such a sustained-release system is a semi-permeable matrix of solidhydrophobic polymers. In certain embodiments, sustained-release systemsmay, depending on their chemical nature, release pharmaceutical agentsover a period of hours, days, weeks or months.

Appropriate pharmaceutical compositions of the present disclosure can bedetermined according to any clinically-acceptable route ofadministration of the composition to the subject. The manner in whichthe composition is administered is dependent, in part, upon the causeand/or location. One skilled in the art will recognize the advantages ofcertain routes of administration. The method includes administering aneffective amount of the agent or compound (or composition comprising theagent or compound) to achieve a desired biological response, e.g., anamount effective to alleviate, ameliorate, or prevent, in whole or inpart, a symptom of a condition to be treated, e.g., oncology andneurology disorders. In various aspects, the route of administration issystemic, e.g., oral or by injection. The agents or compounds, orpharmaceutically acceptable salts or derivatives thereof, areadministered orally, nasally, transdermally, pulmonary, inhalationally,buccally, sublingually, intraperintoneally, subcutaneously,intramuscularly, intravenously, rectally, intrapleurally, intrathecally,intraportally, and parenterally. Alternatively or in addition, the routeof administration is local, e.g., topical, intra-tumor and peri-tumor.In some embodiments, the compound is administered orally.

In certain embodiments, a pharmaceutical composition of the presentdisclosure is prepared for oral administration. In certain of suchembodiments, a pharmaceutical composition is formulated by combining oneor more agents and pharmaceutically acceptable carriers. Certain of suchcarriers enable pharmaceutical compositions to be formulated as tablets,pills, dragees, capsules, liquids, gels, syrups, slurries, suspensionsand the like, for oral ingestion by a subject. Suitable excipientsinclude, but are not limited to, fillers, such as sugars, includinglactose, sucrose, mannitol, or sorbitol; cellulose preparations such as,for example, maize starch, wheat starch, rice starch, potato starch,gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP). In certain embodiments, such a mixture isoptionally ground and auxiliaries are optionally added. In certainembodiments, pharmaceutical compositions are formed to obtain tablets ordragee cores. In certain embodiments, disintegrating agents (e.g.,cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a saltthereof, such as sodium alginate) are added.

In certain embodiments, dragee cores are provided with coatings. Incertain such embodiments, concentrated sugar solutions may be used,which may optionally contain gum arabic, talc, polyvinyl pyrrolidone,carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquersolutions, and suitable organic solvents or solvent mixtures. Dyestuffsor pigments may be added to tablets or dragee coatings.

In certain embodiments, pharmaceutical compositions for oraladministration are push-fit capsules made of gelatin. Certain of suchpush-fit capsules comprise one or more pharmaceutical agents of thepresent invention in admixture with one or more filler such as lactose,binders such as starches, and/or lubricants such as talc or magnesiumstearate and, optionally, stabilizers. In certain embodiments,pharmaceutical compositions for oral administration are soft, sealedcapsules made of gelatin and a plasticizer, such as glycerol orsorbitol. In certain soft capsules, one or more pharmaceutical agents ofthe present invention are be dissolved or suspended in suitable liquids,such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers may be added.

In certain embodiments, pharmaceutical compositions are prepared forbuccal administration. Certain of such pharmaceutical compositions aretablets or lozenges formulated in conventional manner.

In certain embodiments, a pharmaceutical composition is prepared fortransmucosal administration. In certain of such embodiments penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art.

In certain embodiments, a pharmaceutical composition is prepared foradministration by inhalation. Certain of such pharmaceuticalcompositions for inhalation are prepared in the form of an aerosol sprayin a pressurized pack or a nebulizer. Certain of such pharmaceuticalcompositions comprise a propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In certain embodiments using a pressurized aerosol,the dosage unit may be determined with a valve that delivers a meteredamount. In certain embodiments, capsules and cartridges for use in aninhaler or insufflator may be formulated. Certain of such formulationscomprise a powder mixture of a pharmaceutical agent of the invention anda suitable powder base such as lactose or starch.

In other embodiments the compound of the present disclosure areadministered by the intravenous route. In further embodiments, theparenteral administration may be provided in a bolus or by infusion.

In certain embodiments, a pharmaceutical composition is prepared forrectal administration, such as a suppository or retention enema. Certainof such pharmaceutical compositions comprise known ingredients, such ascocoa butter and/or other glycerides.

In certain embodiments, a pharmaceutical composition is prepared fortopical administration. Certain of such pharmaceutical compositionscomprise bland moisturizing bases, such as ointments or creams.Exemplary suitable ointment bases include, but are not limited to,petrolatum, petrolatum plus volatile silicones, and lanolin and water inoil emulsions. Exemplary suitable cream bases include, but are notlimited to, cold cream and hydrophilic ointment.

In certain embodiments, the therapeutically effective amount issufficient to prevent, alleviate or ameliorate symptoms of a disease orto prolong the survival of the subject being treated. Determination of atherapeutically effective amount is well within the capability of thoseskilled in the art.

In certain embodiments, one or more compounds of formula (1) (I), (I′),(I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or apharmaceutically acceptable salt or solvate thereof are formulated as aprodrug. In certain embodiments, upon in vivo administration, a prodrugis chemically converted to the biologically, pharmaceutically ortherapeutically more active form. In certain embodiments, prodrugs areuseful because they are easier to administer than the correspondingactive form. For example, in certain instances, a prodrug may be morebioavailable (e.g., through oral administration) than is thecorresponding active form. In certain instances, a prodrug may haveimproved solubility compared to the corresponding active form. Incertain embodiments, prodrugs are less water soluble than thecorresponding active form. In certain instances, such prodrugs possesssuperior transmittal across cell membranes, where water solubility isdetrimental to mobility. In certain embodiments, a prodrug is an ester.In certain such embodiments, the ester is metabolically hydrolyzed tocarboxylic acid upon administration. In certain instances the carboxylicacid containing compound is the corresponding active form. In certainembodiments, a prodrug comprises a short peptide (polyaminoacid) boundto an acid group. In certain of such embodiments, the peptide is cleavedupon administration to form the corresponding active form.

In certain embodiments, a prodrug is produced by modifying apharmaceutically active compound such that the active compound will beregenerated upon in vivo administration. The prodrug can be designed toalter the metabolic stability or the transport characteristics of adrug, to mask side effects or toxicity, to improve the flavor of a drugor to alter other characteristics or properties of a drug. By virtue ofknowledge of pharmacodynamic processes and drug metabolism in vivo,those of skill in this art, once a pharmaceutically active compound isknown, can design prodrugs of the compound (see, e.g., Nogrady (1985)Medicinal Chemistry A Biochemical Approach, Oxford University Press, NewYork, pages 388-392).

In various aspects, the amount of the compound of formula (1) (I), (I′),(I″), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (2), or (II), or apharmaceutically acceptable salt or solvate thereof, or compoundsdisclosed in Tables 1, 2, and/or Table 3, or a pharmaceuticallyacceptable salt or solvate thereof, can be administered at about 0.001mg/kg to about 100 mg/kg body weight (e.g., about 0.01 mg/kg to about 10mg/kg or about 0.1 mg/kg to about 5 mg/kg).

The concentration of a disclosed compound in a pharmaceuticallyacceptable mixture will vary depending on several factors, including thedosage of the compound to be administered, the pharmacokineticcharacteristics of the compound(s) employed, and the route ofadministration. The agent may be administered in a single dose or inrepeat doses. The dosage regimen utilizing the compounds of the presentinvention is selected in accordance with a variety of factors includingtype, species, age, weight, sex and medical condition of the patient;the severity of the condition to be treated; the route ofadministration; the renal and hepatic function of the patient; and theparticular compound or salt thereof employed. Treatments may beadministered daily or more frequently depending upon a number offactors, including the overall health of a patient, and the formulationand route of administration of the selected compound(s). An ordinarilyskilled physician or veterinarian can readily determine and prescribethe effective amount of the drug required to prevent, counter or arrestthe progress of the condition.

The compounds or pharmaceutical compositions of the present disclosuremay be manufactured and/or administered in single or multiple unit doseforms.

Having now generally described the invention, the same will be morereadily understood through reference to the following examples, whichare provided by way of illustration and are not intended to be limitingof the present invention.

EXAMPLES Example 1 Identification of Parkin Activators

Assay Principle:

The assay based on the irreversible reaction of an Activity-Based Probe(ABP) with the active site cysteine in the enzyme. ABP consists of aubiquitin moiety with an epitope tag (e.g. HA tag) at the N-terminus,and a reactive group at the C-terminus. The activity of Parkin-RBR (w/othe R0 inhibitory domain) is significantly higher than the activity ofParkin-R0RBR or the activity of full-length Parkin. The covalentattachment of ABP to Parkin can be monitored by Time ResolvedFluorescence Resonance Energy Transfer (TR-FRET)

-   -   Parkin-R0RBR, full-length Parkin→low TR-FRET signal (negative        control)    -   Parkin RBR→high TR-FRET signal (positive control)

Compounds increasing the activity of Parkin-R0RBR or the activity offull-length—Parkin can be identified by an increase in TR-FRET signal.

Strategy: use of N-terminal His-SUMO tagged constructs of Parkin-R0RBR,full-length Parkin and Parkin-RBR. (from Evotec Slides; Based on Rileyet al. 2013. Nat Commun. 4:1982 & on information provided by E3× Bio;grant Application)

Constructs:

-   -   Full-length Parkin (1-465), R0RBR (141-465) and RBR (238-465)        expression with N-terminal His₆-SUMO-tag (can potentially be        removed during purification using SENP1 protease) in E. coli as        described by Riley et al.    -   N-terminal His₆-tag enabling TR-FRET-assay→use of the purified        protein that still have the N-terminal His₆-SUMO-tags on.    -   Small scale tests are conducted for all constructs to evaluate        which construct, full-length Parkin or R0RBR, give better yield        to facilitate an HTS-assay.        Phase 1: Protein Production    -   Initiate gene synthesis through third party for full-length        Parkin with N-terminal His₆-SUMO, His₆-SUMO-R0RBR and        His₆-SUMO-RBR, codon-optimized for expression in E. coli and        subcloning into a suitable expression vector    -   Small scale test expression evaluated by Western Blotting to        estimate the yield of soluble protein    -   Transform the RBR construct as well as either the full-length        Parkin construct or the R0RBR construct into BL21(DE3) and        express as outlined in Riley et al., in the scale of 6-24L        (depending on outcome of small scale test expression)    -   Purification of ˜10 mg of the RBR construct as well as either        the full-length Parkin construct or the R0RBR construct as        described by Riley et al.*, i.e. IMAC, MonoQ and size exclusion.        Phase 2: Assay Development

Goals:

-   -   Set-up robust primary screening assays in 1,536-well assay plate        format    -   Establish assays in 384-well format with a reasonable dynamic        range (e.g. using Parkin +/− the R0 inhibitory domain)    -   Optimize assay (e.g. in terms of concentrations of assay        components, buffer, additives, order of addition of reagents,        and incubation temperature)    -   Run time course experiments to define optimal incubation times    -   Demonstrate assay robustness (goal: Z′>0.5)    -   Demonstrate readout stability    -   Test DMSO tolerance    -   Demonstrate specificity of the assay signal obtained using the        Parkin RBR domain (w/o the R0 inhibitory domain) by titration of        Ub (competing with ABP)    -   Transfer assay from 384- to final 1,536-well screening plate        format; adapt the assay to the EVOscreen™ Mark III HTS platform    -   If necessary, fine-tune the assay conditions in order to        optimize assay robustness in this high density plate format        (goal: Z′>0.5) and to demonstrate assay suitability for        high-throughput screening (HTS)    -   Confirm stability of assay reagents under screening conditions        over time    -   Demonstrate plate-to-plate and day-to-day assay robustness    -   Estimate and procure the amounts of all assay reagents required        for screening and hit profiling.        Phase 3: Screening

Marker Library Screen (MLS):

-   -   Pre-screening of a diverse marker library of approximately 2.5 k        representative lead-like compounds against the primary screening        assay at two concentrations in triplicate    -   Statistical analysis of the MLS and hit definition using the        3-sigma-method (plate-based, based on the scatter of        compound-free DMSO wells)    -   Selection of the optimal compound concentration for primary        screening        Primary Screen (PS):    -   Screening of approximately 75,000 lead-like compounds against        the primary screening assay at one uniform compound        concentration (n=1); re-screening of compound plates that do not        meet an agreed re-screen criterion (e.g. Z′>0.5)    -   Hit definition for the primary screen using the 3-sigma-method        (plate-based, based on the scatter of compound-free DMSO wells)    -   Statistical analysis of the primary screen→Primary Hit Compounds        (Parkin activators) Hit Confirmation (HC):    -   Selection of a set of up to approximately 750 primary hits for        Hit Confirmation    -   Cherry picking of the selected compounds and reformatting for        testing    -   Retesting of the selected cpds against the primary screening        assay at the compound screening concentration (n=3)    -   Statistical analysis of the Hit Confirmation        campaign→Identification of confirmed small molecule Parkin        activators.        Phase 4: HitProfiling (HP):    -   Selection of a set of up to approximately 250 confirmed hit        compounds for Hit Profiling    -   Cherry picking of the selected compounds and reformatting for        concentration-response testing    -   Concentration-response testing as 11-point compound dilution        series against the primary screening assay (n=2)    -   Automated data fitting of the concentration response curves and        calculation of the resulting IC50 values    -   LC/MS inspection of the hit compounds to confirm compound        identity and purity    -   Structure-activity relationship analysis (SAR) of the active hit        compounds    -   Confirmed & profiled small molecule Parkin activators.

Example 2 Activity-Based Probe Assay Using an Ubiquitin Vinyl SulfoneProbe

An Ubiquitin vinyl sulfone probe can be used that irreversibly binds tothe active site cysteine of Parkin ligase. Covalent attachment of theprobe to the Parkin can be monitored by TR-FRET. Candidate activatorcompounds can be identified by increasing the activity of Parkin ligasedue to an increase in TR-FRET signal. Screening for activating compoundscan be distinguished from the controls as follows:

-   100% activation signal=Heat activated Parkin+100 nM control    activator in DMSO.-   0% activation signal=Heat activated Parkin+DMSO only.-   Parkin activators can be identified by an increase of the 0%    activation signal TR-FRET signal.

Assay Conditions:

Materials:

-   Assay Plate: White 384 well plate (Corning 3572)-   Enzyme: Parkin-His tagged 203 μM (10.5 mg/ml)-   Probe: Ubiquitin vinyl-sulfone (HA-Ub-VS Boston Biochem U-212)-   DMSO: DMSO (Sigma cat # D4540-100ML)-   Reaction Buffer: 50 mM HEPES (pH 8.5), 150 mM NaCl, 0.01% Tween 20,    0.1% BSA-   Detection Buffer: 50 mM HEPES (pH 8.5), 150 mM NaCl, 0.01% Tween 20,    0.1% BSA, 800 mM KF-   Detection Reagent A: 2.6 nM Anti-6HIS-Eu cryptate and 40 nM    Anti-HA-XL665 in detection buffer-   Eu cryptate: Anti-6HIS-Eu cryptate (CisBio 61HISKLA)-   XL665: Anti-HA-XL665 (CisBio 610HAXLA)    Enzyme Reaction (15 min pre incubation Parkin with activator only)-   Parkin: 40 nM-   HA-Ub-VS Probe: 70 nM-   Activator/DMSO: 2× Activator/2% DMSO-   Reaction time: 60 minutes-   Temperature: 22° C.-   Total volume: 10 μl reaction    Detection Reaction-   Take 10 μl of Enzyme Reaction above and add 10 μl detection Reagent    A under the following conditions:-   Reaction time: 60 minutes-   Temperature: 22° C.-   Total volume: 20 μl

Assay procedure (Using HP D-300 compound dispenser and Bravo for theoperation):

-   1) Heat activate Parkin in reaction buffer (500 μl/1.5 ml tube:    Eppendorf Thermomixer 5 minutes, 400 rpm at 58° C. and put on ice    until needed).-   2) Load assay plate wells with 4.8 μl 84.5 nM Parkin in reaction    buffer by use of Bravo.-   3) Deliver 0.2 μl 200× activator candidates in DMSO by use of HP    D-300 compound dispenser. Highest 200× concentration=20 m and then    twofold dilutions.-   4) Spin 1000 rpm, 2 minutes, at room temp.-   5) Incubate plate for 15 minutes at room temp.-   6) Add 5 μl 140 nM HA-Ub-VS Probe in reaction buffer by use of    Bravo.-   7) Spin 1000 rpm, 2 minutes, at room temp.-   8) Incubate plate for 60 minutes at room temp.-   9) Add 10 μl 2.6 nM Anti-6HIS-Eu cryptate and 40 nM Anti-HA-XL665 in    detection buffer.-   10) Spin 1000 rpm, 2 minutes, at room temp.-   11) Incubate plate for 60 minutes at room temp.-   12) Read plates on Perkin Elmer Envision instrument with the    following parameters:    -   LANCE dual laser protocol loaded into the Envision® software    -   Top Mirror: LANCE/DELFIA Duel/Bias (Bar code 446)    -   Emission Filter: APC 665 EM (Bar code 205)    -   2nd Emission Filter: Europium 615 EM (Bar code 203)    -   Read 655 nm (channel 1) and 615 nm (channel 2) wavelengths on        Envision®

Data Analysis: The Data can be read in CSV files. There are two tablesin those CSV files, which are the values of 655 nm (channel 1) and 615nm (channel 2) wavelengths respectively. The data is converted to anHTRF Ratio=(Channel 1/Channel 2)*10,000

The average of all the 0 uM controls (DMSO only)=BKGD (Background—0%activation). Subtract BKGD from each HTRF Ratio value=HTRF-BKGD. Theaverage of all the 100 uM 100 nM control activator in DMSO controls=Max(100% activation). The following equation is then used to calculate %Activation for each well/candidate as follows: %Activation=(HTRF-BKGD/Max)*100.

The % Activation of compound titration can then be used to findactivation EC50 or highest % activation if less than 75% activation isseen for the candidate compound.

Graphpad Prisim was used with Transform X values: X=Log(X) and nonlinearregression (dose-response-stimulation): Log(agonist) vsResponse—variable slope (four parameters) with constrains set toBottom=0 and Top=100.

The Activity-Based Probe Assay was performed with various compounds inTables 1-3. As shown in Table 4, the compounds indicated a range ofincreasing Parkin activity with the activity-based probe Ubiquitin-vinylsulfone. This is also demonstrated in FIGS. 1 and 4, compoundsN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide andN-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide, respectively.

TABLE 4 Probe Assay Auto-ubiq Compound EC50 (μM) EC50 (μM) Cell RatingsID [Example 2] [Example 3] [Example 4] A 7.0 2.3 +++ B >100 >40 NA C15.10 >40 +++ +++ D >100 >40 NA E 15.30 >40 NA F 6.70 >40 +++ G >100 >40NA H >100 >40 NA I >100 >40 NA J >100 >40 NA K 10.80 >40 +++ L 1.80 >40++ M >100 >40 NA T >100 >40 NA U >100 >40 NA V >100 >40 NA W >100 >40 NAX >100 >40 NA Y >100 >40 NA Z 9.1 >40 + A1 >100 >40 NA B1 >100 >40 NA C13.7 >100 + D1 38.9 >40 NA E1 >100 >40 NA F1 >100 >40 NA G1 >100 >40 NAH1 >100 >40 NA I1 >100 >40 NA J1 >100 >40 NA K1 9.2 >40 NA L1 37.00 >40NA M1 11.9 >40 NA N1 >100 >40 NA O1 12.8 >40 NA T1 >100 >40 NA U125.9 >40 NA V1 >100 >40 NA W1 10.4 >40 NA X1 62.0 >40 NA Y1 2.0 >40 +++Z1 1.0 >40 NA A2 50 >40 NA B2 4.0 >40 NA C2 4.0 >40 NA D2 89.0 >40 NA E26.0 >40 NA F2 12.0 >40 NA G2 2.0 >40 NA H2 >100 >40 NA I2 >100 >40 NA J26.0 >40 NA K2 6.0 >40 NA L2 >100 >40 NA M2 >100 >40 NA N2 >100 >40 NAO2 >100 >40 NA P2 >100 >40 NA Q2 26.0 >40 NA R2 4.0 >40 NA S2 >100 >40NA T2 >100 >40 NA U2 22.0 >40 NA V2 >100 >40 NA W2 >100 >40 NAX2 >100 >40 NA Y2 >100 NA NA A3 >100 NA NA B3 >100 NA NA C3 >100 NA NAD3 >100 NA NA E3 >100 NA NA F3 >100 NA NA G3 74.0 NA NA H3 3.0 NA NAI3 >100 NA NA J3 37.0 NA NA K3 17.0 NA NA L3 >100 NA NA M3 >100 NA NAN3 >100 NA NA O3 >100 NA NA P3 9.0 NA NA Q3 4.0 NA NA R3 3.0 NA NA S33.0 NA NA +++ >70% effect at 10 μM; ++ 69%-31% effect at 10 μM; + <30%effect at 10 μM; NA = not available

Example 3 Parkin pUB Auto-ubiquitinylation Assay

A Parkin pUB Auto-ubiquitinylation Assay is used to evaluate acompound's potency to activate Parkin's ability to Auto-ubiquitinylateitself.

The principle of this assay is that the E3 Ligase Parkin catalyzes thetransfer of Ubiquitin to target proteins, but also has the ability toauto-ubiquitinylate itself. The phospho-Ubiquition (pUb) added to theassay alters the Parkin to a state where small molecule activators canenable the Parkin to auto-ubiquitinylate though the E1-E2 cascadereaction. The use of a Eu cryptate Ubiquition and anti 6His-d2 thatbinds to the His tagged Parkin will give a signal when the Eu cryptateUbiquition is auto-ubiquitinylate onto the Parkin which can be monitoredby TR-FRET.

Similar to the Activity-based probe assay in Example 2, screening foractivating compounds can be distinguished from the controls as follows:

-   100% activation signal=pUb activated Parkin+40 nM control activator    in DMSO.-   0% activation signal=pUb activated Parkin+DMSO only.-   Parkin activators can be identified by an increase of the 0%    activation signal TR-FRET signal.    Materials:-   Assay Plate: White 384 well plate (Corning 3572)-   Enzyme 1: E1 (Ubiquitin-activating enzyme/UBE1 Boston Biochem E-305)-   Enzyme 2: E2 (UBcH7/Ube2L3 Boston Biochem E2-640)-   Enzyme 3: Parkin-His tagged 203 μM (10.5 mg/ml)-   pUb: Phospho-Ubiquitin (S65) (Boston Biochem U-102)-   Eu Cryptate Reagent: Ubiquitin Eu (CisBio 61UBIKLA)-   DMSO: DMSO (Sigma-34869-2.5L)-   Reaction Buffer: 50 mM HEPES, 50 mM NaCl, 1 mM MgCl₂, 0.005% Tween    20, 0.1% PF-127 (Fisher Scientific 50-310-494), pH 8.5-   Detection Buffer: 50 mM HEPES, 50 mM NaCl, 800 mM KF, 5 mM EDTA,    0.005% Tween 20, 0.1% PF-127, pH 8.5-   Detection Reagent Z: 13.4 nM Anti-6His-d2 in detection buffer-   d2 Reagent: Anti-6His-d2 (CisBio 61HISDLA)

Assay Conditions:

Enzyme Reaction (15 min pre-incubation with Parkin, pUb and activatoronly)

-   Parkin: 196 nM-   pUb: 196 nM-   DMSO: 1% DMSO-   E1: 5 nM-   E2: 50 nM-   Ubiquitin Eu: 8.8 nM-   Reaction time: 120 minutes-   Temperature: 22° C.-   Total volume: 10 μl reaction

Detection Reaction

Take 10 μl of Enzyme Reaction above and add 10 μl detection Reagent Zunder the following conditions:

Reaction time: 60 minutes

Temperature: 22° C.

Total volume: 20 μl

Assay Procedure:

-   1) Load assay plate wells with 4.9 μl 400.0 nM Parkin, 400 nM pUb in    reaction buffer by use of Eppendorf 12-channel pipette.-   2) Deliver 0.1 μl 100× activator candidates in DMSO by use of Echo    555 compound dispenser. Highest 100× concentration=100 μm and then    twofold dilutions. Add each compound and control in duplicate wells.-   3) Spin 1000 rpm, 2 minutes, at room temp.-   4) Incubate plate for 15 minutes at room temp.-   5) Add 5 μl 10 nM E1, 100 nM E2, 17.6 nM Ubiquitin Eu and 2 mM ATP    in Reaction Buffer by use of Eppendorf 12-channel pipette.-   6) Spin 1000 rpm, 2 minutes, at room temp.-   7) Incubate plate for 120 minutes at room temp.-   8) Add 10 μl 13.4 nM anti his d2 in detection buffer by use of    Eppendorf 12-channel pipette.-   9) Spin 1000 rpm, 2 minutes, at room temp.-   10) Incubate plate for 120 minutes at room temp.-   11) Read plates on Perkin Elmer Envision instrument with the    following parameters:    -   LANCE dual laser protocol loaded into the Envision® software    -   Top Mirror: LANCE/DELFIA Duel/Bias (Bar code 446)    -   Emission Filter: APC 665 EM (Bar code 205)    -   2nd Emission Filter: Europium 615 EM (Bar code 203)    -   Read 655 nm (channel 1) and 615 nm (channel 2) wavelengths on        Envision®

Data Analysis: The Data can be read in CSV files. There are two tablesin those CSV files, which are the values of 655 nm (channel 1) and 615nm (channel 2) wavelengths respectively. The data is converted to anHTRF Ratio=(Channel 1/Channel 2)*10,000

The average of all the 0 uM controls (DMSO only)=BKGD (Background—0%activation). Subtract BKGD from each HTRF Ratio value=HTRF-BKGD. Theaverage of all the 100 uM control activator in DMSO controls=Max (100%activation). The following equation is then used to calculate %Activation for each well/candidate as follows: %Activation=(HTRF-BKGD/Max)*100.

The % Activation of compound titration can then be used to findactivation EC50 or highest % activation if less than 75% activation isseen for the candidate compound.

XLFIT5 model 205 was applied for the data analysis. EC50 fit model (4Parameter Logistic Model/Sigmoidal dose-Response Model);fit=(A+((B−A)/(1+((C/x)^D)))); res=(y-fit). The parameters are:

A: Bottom

B: Top

C: Relative EC50

D: Hill Slope

Constrains set to Bottom=0 and Top=100.

This Parkin pUB auto-ubiquitinylation Assay was performed with variouscompounds in Tables 1, 2, and/or Table 3. The compounds indicated rangeof increasing Parkin activity in an auto-ubiquitination assay as shownin Table 4. This is also demonstrated in FIGS. 2 and 5 for compoundsN,N′-(1-phenyl-1H-1,2,4-triazole-3,5-diyl)dibenzamide andN-(5-cinnamamido-1-phenyl-1H-1,2,4-triazol-3-yl)benzamide.

Example 4 Cell Rating Experiments

Compounds: All compounds were dissolved in DMSO to a concentration of 25mM and stored at −20° C.

Cell Culture: S-HeLa stably expressing a YFP-Parkin fusion protein(kindly donated by Prof Richard J. Youle, Porter Neuroscience ResearchCenter, Bethesda, Md., USA) were utilised to assess Parkin-dependentinduction of mitophagy. 4000 cells were seeded in each well of a 96 wellplate (Parkin Elmer ViewPlate-96 F TC, cat. N. 6005182) and left to growfor 24 hours.

Subsequently cells were incubated with vehicle (DMSO) or 6 μM CCCP for 1hour prior to adding increasing concentrations of compound (1, 2.5, 5,10 μM), each condition run in replicate of five. After 20 hours cellswere processed for immunofluorescence.

Immunofluorescence: Samples were fixed in 4% PFA for 25 minutes RT andpermeabilized with PBS 0.1% Triton-X100 for 3 minutes on ice, blockedwith PBS 3% BSA, 0.3% Triton-X100 for 2 hours RT, followed by overnightincubation with primary antibody at 4° C. (0.5 μg/ml rabbit Tomm20antibody FL-145; Santa Cruz Biotechnology) diluted in PBS 0.1% BSA, 0.3%Triton-X100. The secondary goat anti-rabbit antibody conjugated withDyLight 649 (Jackson ImmunoResearch) was applied for 1 hour at roomtemperature at a concentration of 2.8 μg/ml in conjunction with 1 μg/mlHoechst33342.

Cells were imaged using an Olympus ScanR automated microscope equippedwith motorised stage and 20×APO planar objective. 18 images wereacquired for each well using the following combination ofexcitation/emission filters: Hoechst33342 was excited through a 350/50nm band pass filter and fluorescence intensity was collected through a460/30 band pass filter. YFP was excited through a 500/20 nm band passfilter and fluorescence intensity was collected through a 540/35 bandpass filter. DyLight 649 was excited through a 640/30 nm band passfilter and fluorescence intensity was collected through 700/75 band passfilter. Images were processed and analysed as described in the ImageAnalysis section.

Image Analysis: Images were processed and analysed using Columbus HCSAnalysis software (Version 2.5.0., PerkinElmer) as follows:

Tomm20 fluorescence intensity was corrected using the parabolaalgorithm. Hoechst 33342 fluorescence was used to identify and countcells. Cells were segmented according to Tomm20 fluorescence intensity.Spot detection was optimized to recognize number and total cellular areaof Tomm20 stained clusters (mitochondria).

Tomm20 staining intensity, spot numbers and spot area were used to traina linear classifier algorithm that discriminated between Tomm20 positive(high intensity, spot numbers and spot area) and Tomm20 negative cells(low intensity, spot numbers and spot area).

Bar graphs were generated reporting the number of Tomm20 negative cellsexpressed as percentage of total cells imaged for each well (FIGS. 3 and6). Results were shown as mean±SD of a representative experimentperformed in triplicate. +++ indicates >70% effect at 10 μM; ++indicates 69%-31% effect at 10 μM; + indicates <30% effect at 10 μM;NA=not available.

Example 5 Microsomal Stability Assay

Compounds were also tested for metabolic stability in both rat livermicrosomes (RLM) and human liver microsomes (HLM) and their half-lifecalculated (See Table 5). The assay was performed as follows. The totalvolume for each incubation was 250 μL. A 100 μM DMSO solution ofcompound (diluted from 10 mM stock solution) was spiked into 50 mMKH₂PO₄ (pH 7.4) buffer containing liver microsome at a concentration of1.0 mg/mL. The reaction was initiated by the addition of 50 μL of 1 mMNADPH. The final concentration of each compound was 1 μM (1% DMSO). Thepositive controls, phenacetin for CYP1A2, diclofenac for CYP2C9,omeprazole for CYP2C19, dextromethorphan for CYP2D6 and midazolam forCYP3A4 were added to a separate tube with the final substrateconcentrations of 1 μM (1% DMSO) for evaluating the enzyme activities inthe liver microsomes. At 0, 15, 30 and 60 min, an aliquot of 15 μLreaction mixtures were removed and 200 μL of methanol (with internalstandard of 25 ng/mL propranolol) was added to quench the reaction. Theresulting mixture was centrifuged and supernatant was used for LC-MS/MSanalysis.

The signals for each compound, or the metabolites for the probesubstrates and the internal standard were integrated and the peak arearatios to internal standard were generated. Percent parent remaining ata specified timepoint was calculated based on the peak area ratios attime 0 (as 100%) for in vitro metabolic stability studies in livermicrosome and hepatocyte. The observed rate constant (k_(obs)) for themetabolism of substrates was calculated by plotting the natural log ofpercentage substrate remaining versus time of incubation with the slopebeing k_(obs). The half-life (T_(1/2)) was calculated according to thefollowing equation:T _(1/2)=0.693/k _(obs)

A long half-life, such as with Compound F, suggests metabolic stabilityin the liver, whereas a short half-life, such as Compound E2, suggests ahigh susceptibility to metabolism in the liver.

TABLE 5 Compound HLM t_(1/2) RLM t_(1/2) ID (min) (min) A 151.0 20.6 C77.9 14.1 F 578.0 630.0 L 38.5 59.2 Z NA 45.0 U1 6.9 10.5 W1 NA 110.0 Y1NA <1 Z1 NA 19.0 A2 NA 36.0 B2 NA 2.0 C2 NA 12.0 E2 NA 3.0 G2 NA 39.0 J2NA 50.0 K2 NA 29.0 Q2 NA 76.0 R2 NA 77.0 U2 NA 126.0 H3 NA 16.0 Q3 NA133

Example 6 Oral Bioavailability (PO) and Pharmacokinetics Assay

The pharmacokinetics and oral bioavailability of various compounds wasevaluated following IV or PO administration to fasted maleSprague-Dawley rats (N=3/route/dose). Each compound was administered tofasted male Sprague-Dawley rats as a single dose of 1 mg/kg (IV) or 5mg/kg (PO). For IV dosing, each compound was formulated in 10% DMA/15%Solutol/75% HPβ-CD (20%) as a 1 mg/mL solution for IV (1 mL/kg) and forPO (5 mL/kg) administrations.

Blood samples (0.250 mL) were collected into EDTA tubes then processedto generate plasma samples. Blood samples were collected at pre-dose,0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hours post-dose administration.Plasma concentrations of each compound were determined using LC-MS/MSwith a lower limit of quantitation of 1.0 ng/mL. The pharmacokineticparameters were determined by non-compartmental methods using WinNonlin.

The pharmacokinetic parameters of each compound were determined bynon-compartmental analysis (Model 201 for IV administration and Model200 for PO administration) using WinNonlin Version 6.4 (Pharsight,Mountain View, Calif.). The area under the curve from the time of dosingto the last measurable concentration, AUC_((0-t)), was calculated by thelinear trapezoidal rule. The area under the concentration-time curveextrapolated to infinity, AUC_((0-∞)), was calculated as follows:AUC_((0-∞))=AUC_((0-t))+C_(last)/k

Where C_(last) is the last measurable concentration and k is the firstorder rate constant associated with the terminal elimination phase,estimated by linear regression of log concentration versus time. Thehalf-life (T_(1/2)) of the terminal elimination phase was estimatedbased on the following equation: T_(1/2)=0.693/k

K was determined based on at least three timepoints with R²≥0.9.Additional parameters were calculated as follows: CL=Dose/AUC_((0-∞))

Where CL is the clearance of An2H compound in L/hr/kg, Dose is theadministered dose in mg/kg. Mean residence time (MRT) was calculated asfollows: MRT=AUMC_((0-∞))/AUC_((0-∞))

Where area under the first moment curve extrapolated to infinity(AUMC_((0-∞))) was calculated as follows:AUMC_((0-∞))=AUMClast+tlast*Clast/k+Clast/k²

The steady state volume of distribution (Vss) was calculated as follows:Vss=CL×MRT.

The oral bioavailability based on AUC_((0-t)) was calculated as follows:F (%)=AUC_(PO)/AUC_(IV)×Dose_(IV)/Dose_(PO)×100. For IV administration,the initial concentration, Co is reported and is an extrapolated value.For PO administration, the maximum concentration, Cmax is report and isan observed value.

FIGS. 7-11 show intravenous (IV) and oral (PO) plasma concentrationcurves for Compounds A, F, K, H and C. Table 6 (directly below) showsthe oral bioavailability of Compounds A, F, K, H and C Tables 7-11 showthe PK parameters of Compounds A, F, K, H and C.

TABLE 6 Oral Bioavailability Study Oral Compound Bioavilability ID (POPK) A F = 7% C F = 2% F F = 24% H F = 6% K F = 4%

TABLE 7 Rat Intravenous (IV) and oral (PO) bioavailability for CompoundA Route IV bolus (dose) (1 mg/kg) PO (5 mg/kg) Formulation PEG 400/HPβCDC0 or Cmax (ng/mL) 4520 249 Tmax (hr) NA 0.417 T½ (hr) 0.85 1.62 AUC0-t(ng · hr/mL) 1099 371 Cl (mL/min/kg) 15.7 NA Vss (L/kg) 0.381 NA F (%)100 6.85

Compound A had an oral bioavailability of 7% in rats (Table 6) withT_(1/2) of 1.6 hours (Table 7). Compound A had medium clearance by IVadministration in rats with 15.7 mL/min/kg and Vss (steady state volumeof distribution) of 0.38 L/kg and T_(1/2) of 0.85 hours (Table 7).

TABLE 8 Rat Intravenous (IV) and Intraperitoneal (IP) bioavailabilityfor Compound F Route IV bolus (dose) (1 mg/kg) IP (3 mg/kg) Formulation10% DMA/15% solutol/ 75% HP-b-CD (20%) C0 or Cmax (ng/mL) 5981 1467 Tmax(hr) NA 1.3 T½ (hr) 2.5 3.2 AUC0-t (ng · hr/mL) 4274 9589 Cl (mL/min/kg)4.03 NA Vss (L/kg) 0.406 NA In vitro RLM (min) Stable (F: 75%)Compound F had low IV clearance and small Vss with T_(1/2) of 2.5 hours,consistent with in vitro RLM stability (Table 8). Compound F had highsystemic exposures following IP injection with systemic bioavailabilityof 75% (Table 8). Compound F further had a high oral bioavailability(24%) in rats relative to compounds of similar structure (Table 6).

TABLE 9 Rat Intravenous (IV) and oral (PO) bioavailability for CompoundK Route IV bolus (dose) (1 mg/kg) PO (5 mg/kg) Formulation 10% DMA/15%solutol/ 75% HP-b-CD (20%) C0 or Cmax 2121 52.4 (ng/mL) Tmax (hr) NA0.42 T½ (hr) 0.60 2.1 AUC0-t (ng · hr/mL) 902 172 Cl (mL/min/kg) 18.4 NAVss (L/kg) 0.707 NA In vitro RLM (min) ND (F: 4.0%)

Compound K had moderate IV clearance and Vss of 0.71 L/kg with T_(1/2)of 0.6 hours (Table 9). Compound K had a low oral bioavailability of4.0% following oral dosing (Tables 6 and 9).

TABLE 10 Rat Intravenous (IV) and oral (PO) bioavailability for CompoundH Route IV bolus (dose) (1 mg/kg) PO (5 mg/kg) Formulation 10% DMA/15%solutol/ 75% HP-b-CD (20%) C0 or Cmax 6736 91.0 (ng/mL) Tmax (hr) NA1.33 T½ (hr) 0.69 1.7 AUC0-t (ng · hr/mL) 1374 410 Cl (mL/min/kg) 12.2NA Vss (L/kg) 0.401 NA In vitro RLM (min) ND (F: 6.2%)

Compound H had moderate IV clearance and Vss of 0.40 L/kg with T_(1/2)of 0.69 hours (Table 10). Compound H had a low oral bioavailability of6.2% following oral dosing (Tables 6 and 10).

TABLE 11 Rat Intravenous (IV) and oral (PO) bioavailability for CompoundC Route IV bolus (dose) (1 mg/kg) PO (5 mg/kg) Formulation 10% DMA/15%solutol/ 75% HP-b-CD (20%) C0 or Cmax 3985 50.5 (ng/mL) Tmax (hr) NA0.38 T½ (hr) 0.71 2.3 AUC0-t (ng · hr/mL) 1026 114 Cl (mL/min/kg) 16.6NA Vss (L/kg) 0.416 NA In vitro RLM (min) 14.1 (F: 2.4%)

Compound C had moderate IV clearance and Vss of 0.42 L/kg with T_(1/2)of 0.71 hours, not predicted by in vitro RLM stability (potential highprotein binding) (Table 11). Compound C had a low oral bioavailabilityof 2.4% following oral dosing (Tables 6 and 11).

Example 7 In Vivo Cancer Xenograft Assay

Compound F was evaluated for in vivo therapeutic efficacy in thetreatment of subcutaneous HCT-116 (colon cancer) and Calu-6 (lungcancer) xenograft models in nude mice.

Each mouse was inoculated subcutaneously at the flank region with eitherHCT-116 or Calu-6 tumor cells (1.0×106) in 0.1 ml of 1×PBS mixed withMatrigel (1:1) for tumor development and xenotransplantation.

Twenty (20) animals with approx 120-150 mm tumors were selected forHCT-116 follow up experiment and randomly placed into Groups 1, 2, 3,and 4, wherein Group 1 was for mice administered with a vehicle negativecontrol; Group 2 was positive control of Mice treated with Bevacizumabat 5 mg/kg every three days; Group 3 was mice treated with 5 mg/kg ofCompound f daily for up to 20 days; and Group 4 was mice treated with 10mg/kg of Compound F daily for up to 20 days. The vehicle with or withoutdrug was administered to the mouse by intraperitoneal injection. Theformulation with respective drug was as follows:

10%—DMA: N,N-Dimethyl acetamide

15%—Solutol HS 15: Macrogol 15 Hydroxy Stearate, Polyethyleneglycol-15-hydroxystrerate; and 75%-20% aqueous HP-β-CD:Hydroxypropyl-beta-cyclodextrin

Before grouping and treatment, all animals were weighed and the tumorvolumes were confirmed (approx. 120-150 mm3) using electronic caliper.Since the tumor volume can affect the effectiveness of any giventreatment, mice were assigned into groups using randomized block designas following: First, the experimental animals were divided intohomogeneous blocks based on their tumor volume. Secondly, within eachblock, randomization of experimental animals to different groups wereconducted. By using randomized block design to assign experimentalanimals, we ensured that each animal has the same probability of beingassigned to any given treatment groups and therefore systematic errorwas minimized.

After tumor cells inoculation, the animals were checked daily formorbidity and mortality. At the time of routine monitoring, the animalswere checked for any adverse effects of tumor growth and treatments onnormal behavior such as mobility, visual estimation of food and waterconsumption, body weight gain/loss, eye/hair matting and any otherabnormal effects. Death and observed clinical signs were recorded. Tumorvolumes were measured every three days in two dimensions using anelectronic caliper, and the volume data are expressed in mm³ using theformula: V=0.5 a×b2 where a and b are the long and short diameters ofthe tumor, respectively. Tumor volume average was then recorded for eachday for groups 1-4 until day 26. FIG. 12. Shows the xenograft studytesting compound F efficacy to delay subcutaneous HCT-116 tumor growth;FIG. 13 shows the xenograft study testing Compound F efficacy to delaysubcutaneous Calu-6 tumor growth. Compound F was effective of delayinggrowth for both subcutaneous HCT-116 tumor growth and subcutaneousCalu-6 tumor growth.

Example 8 Cancer Cell Proliferation Assay

Measurement of the inhibitive effect of compounds on cancer cellproliferation was performed. Various cell lines were tested, includingHCT-116 (colon); A549 (lung); TOV-21G (ovarian); Calu-6 (lung); H1703(Lung); LS-174T (colon); and SKOV3 (ovarian). The assays were performedunder the following conditions: Cells are harvested at a concentrationof 4×10⁴ cells/ml in media. Volumes of 100 μl/well of these cellsuspensions were added to a 96 well plate using a multichannel pipette.Plates were gently agitated to ensure an even dispersion of cells over agiven plate. Cells were then incubated at 37° C., 5% CO² overnight.Following this, 100 μl of compound at varying concentrations was addedto wells in triplicate. Control wells are those with 100 μl mediacontaining 0.33% DMSO added to cell suspension (this is the equivalentvolume of DMSO found in the highest concentration of drug). Plates werethen gently agitated, as above, and incubated at 37° C., 5% CO₂ for 72hrs (control wells have reached 80-90% confluency). Assessment of cellproliferation in the presence of the compound was determined by the acidphosphatase assay.

Following the incubation period of 72 hrs, media was removed from theplates. Each well on the plate was washed twice with 100 μl PBS. Thiswas then removed and 100 μl of freshly prepared phosphatase substrate(10 mM p-nitrophenol phosphate in 0.1M sodium acetate, 0.1% tritonX-100, pH 5.5) was added to each well. The plates were then incubated inthe dark at 37° C. for 2 hours. Colour development was monitored duringthis time. The enzymatic reaction was stopped by the addition of 50 μlof 1N NaOH. The plate was read in a dual beam plate reader at 405 nmwith a reference wavelength of 620 nm. The absorbance reading of thelatter is subtracted from the former, and the effect of the drug on cellproliferation was then measured as a percentage against the controlcells (DMSO), which is taken as 0% inhibition. Table 12 below shows thatnumerous compounds has nanomolar to micromolar IC₅₀ values on cancercell proliferation including HCT-116 (colon); A549 (lung); TOV-21G(ovarian); Calu-6 (lung); H1703 (Lung); LS-174T (colon); and SKOV3(ovarian). FIG. 14 A-K also show the % inhibition curves of variouscompounds on these cancer cell lines with the 72 hrs proliferationassay.

TABLE 12 IC₅₀ Values of Various Compounds on Cancer Cell LineProliferation C F K L Y1 Z1 Q2 H3 Q3 R3 S3 HCT-116 1.67 uM 2.28 uM 1.92uM  939 nM 351 nM 132 nM 1.64 uM 485 nM 294 nM 1.30 uM 482 nM A549 2.26uM 2.65 uM 2.53 uM 1549 uM  893 nM 253 nM 2.16 uM 823 nM 613 nM 1.98 uM878 nM TOV-21G 1.74 uM 2.22 uM 1.35 uM  989 nM 495 nM 210 nM 1.60 uM 751nM 688 nM 1.19 uM 566 nM Calu6 N/A N/A N/A 2.27 uM 3.34 uM  1.06 uM  N/A2.41 uM  194 nM 3.37 uM 2.36 uM  H1703 2.59 uM N/A 3.46 uM N/A  4.7 uM481 nM ~2.53 uM  3.04 uM  1.54 uM  2.62 uM 2.71 uM  LS-174T 2.20 uM 2.67uM 3.11 uM 1.52 uM 938 nM 391 nM 2.29 uM 497 nM 1.73 uM  1.69 uM 522 nMSKOV3 3.16 uM 3.30 uM 3.14 uM 1.23 uM 1.18 uM  606 nM 2.08 uM 737 nM2.06 uM  2.71 uM 650 nM

Example 9 Synthesis ofN-[5-benzamido-1-(4-iodophenyl)-1,2,4-triazol-3-yl]benzamide (CompoundE)

Step a: Synthesis of (4-iodophenyl)hydrazine

To a suspension of 4-iodoaniline (10.00 g, 45.66 mmol, 1.00 eq) in HCl(12 M, 30 mL) at 0° C. was added dropwise a solution of NaNO₂ (3.15 g,45.66 mmol, 1.00 eq) in H₂O (15 mL), and the resulting mixture wasstirred at 25° C. for 1 h. Then a solution of SnCl₂ (30.91 g, 136.98mmol, 3.00 eq) in HCl (12 M, 20 mL) was added dropwise at 0° C. Thereaction mixture was stirred at 25° C. for 1 h. TLC (DCM/MeOH=10:1)indicated the starting material was consumed completely and a new spotformed. The reaction mixture was filtered and the filter cake was driedunder reduced pressure to afford (4-iodophenyl)hydrazine (9.50 g, HClsalt, crude) as a purplish red solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm:10.39 (br, 3H), 8.50 (br, 1H), 7.59 (d, J=8.8 Hz, 2H), 6.86 (d, J=8.8Hz, 2H).

Step b: Synthesis of 1-(4-iodophenyl)-1,2,4-triazole-3,5-diamine

To a solution of (4-iodophenyl)hydrazine hydrochloride (6.50 g, 24.03mmol, 1.00 eq) in H₂O (15 mL) was added 1-cyanoguanidine (2.02 g, 24.03mmol, 1.00 eq) and HCl (12 M, 5 mL). The reaction mixture was stirred at100° C. for 3 h. TLC (DCM/MeOH=8/1) indicated the starting material wasconsumed completely and new spots formed. The reaction was basified topH=8 with aq. NaOH solution (40%, w/v). After removal of the solventunder vacuum, hexane (100 mL) was added and the resulting mixture wasstirred at 25° C. for 15 min. The mixture was filtered and the filtercake was washed with DCM (150 mL). The solids were collected and driedin vacuo to afford 1-(4-iodophenyl)-1,2,4-triazole-3,5-diamine (1.00 g,crude) as a brown solid. LC-MS (ESI): m/z 301.8 (M+H)⁺.

Step c: Synthesis ofN-[5-benzamido-1-(4-iodophenyl)-1,2,4-triazol-3-yl]benzamide

To a mixture of 1-(4-iodophenyl)-1,2,4-triazole-3,5-diamine (500 mg,1.66 mmol, 1.00 eq) in pyridine (20 mL) was added benzoyl chloride (934mg, 6.64 mmol, 771.69 μL, 4.00 eq), then the mixture was stirred at 100°C. for 5 h. LC-MS indicated the desired product was detected. Ethylacetate (60 mL) was added and the resulting mixture was washed with HCl(1 M, 40 mL×3). The organic layer was dried over anhydrous Na₂SO₄,concentrated to afford the crude product, which was purified byprep-HPLC (column: Phenomenex Synergi C18 150×25×10 μm; mobile phase:[water (0.05% HCl)-ACN]; B %: 50%-70%, 10.5 min) to affordN-[5-benzamido-1-(4-iodophenyl)-1,2,4-triazol-3-yl]benzamide (21.80 mg,42.80 μmol, 3% yield, 99+% purity) as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ ppm: 11.27 (br, 1H), 11.01 (br, 1H), 8.10-8.00 (m, 2H),8.00-7.80 (m, 4H), 7.67-7.60 (m, 2H), 7.56-7.36 (m, 6H); ¹³C NMR (75MHz, DMSO-d₆) δ ppm: 166.8, 165.8, 155.5, 146.1, 138.7, 137.2, 134.0,133.3, 132.6, 129.2, 129.0, 128.4, 125.1, 94.6. LC-MS (ESI): m/z 510.0(M+H)⁺.

Example 10 Synthesis of2-methyl-N-[5-[(2-methylbenzoyl)amino]-1-phenyl-1,2,4-triazol-3-yl]benzamide(Compound F)

Step a: Synthesis of 1-phenyl-1,2,4-triazole-3,5-diamine

To a solution of 1-cyanoguanidine (10.00 g, 118.93 mmol, 1.00 eq) andphenylhydrazine (12.86 g, 118.93 mmol, 11.69 mL, 1.00 eq) in H₂O (40 mL)was added hydrochloric acid (12 M in water, 8 mL). Then the reactionmixture was stirred at 100° C. for 14 h. The reaction mixture wasbasified pH to 8 with 40% sodium hydroxide aqueous solution. Afterremoval of the solvent, the residue was slurried with hexane (150 mL)and followed by DCM (200 mL). The mixture was filtered and the filtercake was collected to give 1-phenyl-1,2,4-triazole-3,5-diamine (36.00 g,197.27 mmol, 83% yield) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ7.48-7.40 (m, 4H), 7.23-7.19 (m, 1H), 6.70 (brs, 2H), 6.23 (brs, 2H).

Step b: Synthesis of2-methyl-N-[5-[(2-methylbenzoyl)amino]-1-phenyl-1,2,4-triazol-3-yl]benzamide

To a solution of 1-phenyl-1,2,4-triazole-3,5-diamine (800 mg, 4.57 mmol,1.00 eq) in pyridine (10 mL) was added 2-methylbenzoyl chloride (2.12 g,13.70 mmol, 1.78 mL, 3.00 eq). The reaction mixture was heated to 110°C. and stirred for 12 h. LC-MS indicated the starting material wasconsumed completely and desired compound was detected. The reactionmixture was quenched by saturated aqueous NH₄Cl (80 mL), and thenextracted with ethyl acetate (100 mL×3). The organic layers werecombined, washed with sat. NH₄C1 (50 mL), and sat.brine (50 mL), thendried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue, which was purified by column chromatographyon silica gel (3-50% ethyl acetate/petroleum ether) to afford the crudeproduct. It was further purified by prep-HPLC (column: PhenomenexSynergi C18 150×25×10μm; mobile phase:[water (0.05% HCl)-ACN]; B %:37%-57%, 10.5 min) to afford2-methyl-N-[5-[(2-methylbenzoyl)amino]-1-phenyl-1,2,4-triazol-3-yl]benzamide(22.0 mg, 52.40 μmol, 98% purity) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ 7.61-7.54 (m, 2H), 7.52-7.50 (m, 4H), 7.42-7.39 (m, 3H),7.31-7.25 (m, 4H), 2.48 (s, 3H), 2.29 (s, 3H); ¹³C NMR (100 MHz, CD₃OD)δ 169.9, 169.6, 154.1, 145.4, 136.7, 136.2, 130.8, 130.7, 129.2, 127.2,127.1, 125.5, 125.5, 125.4, 18.4. LC-MS (ESI): m/z 412.1 (M+H)⁺.

Example 11 Synthesis ofN-(5-benzamido-2-phenyl-1,2,4-triazol-3-yl)-2-methyl-benzamide (CompoundK)

Step a: Synthesis of N-(5-amino-1-phenyl-1,2,4-triazol-3-yl)benzamide

To a mixture of 1-phenyl-1,2,4-triazole-3,5-diamine (1.00 g, 5.71 mmol,1.00 eq), pyridine (490 mg, 6.19 mmol, 500.00 μL, 1.08 eq) in MeCN (5mL) was added dropwise a solution of benzoyl chloride (883 mg, 6.28mmol, 729.69 μL, 1.10 eq) in MeCN (3 mL). The reaction mixture wasstirred at 80° C. for 10 min. LC-MS indicated the desired product wasformed. The mixture was cooled to 10° C. and the mixture was filtered.The filtrate was concentrated to giveN-(5-amino-1-phenyl-1,2,4-triazol-3-yl)benzamide (1.00 g, 2.97 mmol, 52%yield) as a yellow solid. LC-MS (ESI): m/z 280.0 (M+H)⁺.

Step b: Synthesis ofN-(5-benzamido-2-phenyl-1,2,4-triazol-3-yl)-2-methyl-benzamide

A mixture of N-(5-amino-1-phenyl-1,2,4-triazol-3-yl)benzamide (200 mg,716.08 μmol, 1.00 eq), 2-methylbenzoyl chloride (77 mg, 501.26 μmol,65.12 μL, 0.70 eq) in pyridine (3 mL) was stirred at 80° C. for 2 h.LC-MS indicated the desired product was formed. The mixture wasconcentrated to give a crude product, which was purified by prep-HPLC(column: Phenomenex Synergi C18 150×25×10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 35%-55%, 7.8 min) to giveN-(5-benzamido-2-phenyl-1,2,4-triazol-3-yl)-2-methyl-benzamide (67.2 mg,169.09 μmol, 24% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.86(brs, 1H), 7.89 (d, J=7.6 Hz, 2H), 7.77 (d, J=7.6 Hz, 2H), 7.57-7.36 (m,10H), 2.31 (s, 3H). ¹³C NMR (100 MHz, CDCl₃) δ ppm: 164.6, 137.8, 132.7,131.5, 130.1, 128.8, 128.6, 127.9, 127.5, 127.0, 126.8, 17.9. LC-MS(+ESI): m/z 398.2 (M+1)⁺.

Example 12 Synthesis ofN-[1-phenyl-5-(quinazolin-4-ylamino)-1,2,4-triazol-3-yl]benzamide(Compound L)

To a solution of 4-chloroquinazoline (200 mg, 1.23 mmol, 2.00 eq) indioxane (2 mL) was addedN-(5-amino-1-phenyl-1,2,4-triazol-3-yl)benzamide (220 mg, 614.39 μmol,1.00 eq). The mixture was stirred at 150° C. for 2 h using a microwave.LC-MS showed most of the starting material remained but the desiredcompound was detected. The reaction mixture was concentrated undervacuum to give a residue, which was purified by prep-TLC (9%methanol/dichloromethane) to give a crude product. The crude product wasfurther purified by prep-HPLC to affordN-[1-phenyl-5-(quinazolin-4-ylamino)-1,2,4-triazol-3-yl]benzamide (6.2mg, 15.22 μmol, 3% yield) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ11.47 (brs, 1H), 9.00 (br, 1H), 8.16-7.87 (m, 5H), 7.69-7.42 (m, 9H).LC-MS (ESI): m/z 408.1 (M+H)⁺.

Example 13 Synthesis of2-methyl-N-(3-(5-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-yl)benzamide(Compound R3) and2-methyl-N-(3-(3-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-yl)benzamide(Compound S3)

Step A: Synthesis of 3,5-dibromo-1-phenyl-1H-1,2,4-triazole

Two batches of phenylboronic acid (13.5 g, 111 mmol, 1.0 eq),3,5-dibromo-1H-1,2,4-triazole (25 g, 110 mmol, 1.0 eq), Cu(OAc)2 (30 g,165 mmol, 1.5 eq), pyridine (26.5 g, 335 mmol, 27 mL, 3.0 eq) and 4 Å MS(5 g, 22.0 mmol) in toluene (250 mL) was degassed and purged with O2 forthree times, and then the mixture was stirred at 80° C. for 16 h underO2 atmosphere (15 psi). After completion of the reaction, the twobatches of reaction mixture were mixed and filtered, then concentratedunder reduced pressure to give a residue. The residue was purified byflash silica gel chromatography (ISCO®; 200 g SepaFlash® Silica FlashColumn, eluent of 0˜10% ethyl acetate/petroleum ether gradient @ 80mL/min) to give 36 g crude product with 67% purity. 2 g was used fornext step directly. The remaining 34 g was diluted with DCM (200 mL) andwashed with saturated aqueous NaHCO3 (100 mL×1), brine (100 mL), driedover anhydrous Na2SO4, filtered and concentrated under reduced pressureto give 3,5-dibromo-1-phenyl-1,2,4-triazole (29.2 g, 37% yield, 85%purity) as a light yellow solid. LC-MS (ESI): m/z (M+H) 303.9.

Step B: Synthesis of3-bromo-N-(4-methoxybenzyl)-1-phenyl-1H-1,2,4-triazol-5-amine

Two batches of 3,5-dibromo-1-phenyl-1,2,4-triazole (2 g, 5.61 mmol, 1.0eq), (4-methoxyphenyl) methanamine (795 mg, 5.80 mmol, 0.75 mL, 1.0 eq)and K2CO3 (1.16 g, 8.42 mmol, 1.5 eq) in NMP (3 mL) was stirred at 150°C. for 1 h under microwave. TLC (petroleum ether/ethyl acetate=2:1)showed one main spot with desired product was detected for the twobatches. The two batches of reaction mixture were combined, diluted withDCM (80 mL), washed with water (50 mL×3), saturated NaCl (50 mL), driedover Na2SO4, filtered and concentrated under reduced pressure. Theresidue was purified by flash silica gel chromatography (TLC: petroleumether/ethyl acetate=5:1; ISCO®; 20 g SepaFlash® Silica Flash Column,eluent of 0-20% ethyl acetate/petroleum ether gradient @ 80 mL/min) togive 5-bromo-N-[(4-methoxyphenyl)methyl]-2-phenyl-1,2,4-triazol-3-amine(2.4 g, 58% yield, 97% purity) as light yellow oil. LC-MS (ESI): m/z(M+H) 359.1/361.1.

Step C: Synthesis ofN-(4-methoxybenzyl)-3-(5-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-amine

A mixture of5-bromo-N-[(4-methoxyphenyl)methyl]-2-phenyl-1,2,4-triazol-3-amine (500mg, 1.35 mmol, 1.0 eq), tributyl-(5-methyl-2-pyridyl)stannane (500 mg,1.31 mmol, 0.97 eq) and [2-(2-aminophenyl)phenyl]-chloro-palladium;dicyclohexyl-[2-(2,6-dimethoxyphenyl)phenyl]phosphane (100 mg, 139 μmol,0.1 eq) in THF (20 mL) was degassed and purged with N2 for three times,and then the mixture was stirred at 90° C. for 16 h under N2. Most ofstart material remained. Then the reaction mixture was filtered and[2-(2-aminophenyl)phenyl]-chloro-palladium;dicyclohexyl-[2-(2,6-dimethoxyphenyl)phenyl]phosphane (100 mg, 139 μmol,0.1 eq) was added to the mixture. The reaction mixture was stirred at90° C. for 72 h. After being cooled to room temperature, the reactionmixture was quenched by addition of saturated aqueous KF (20 mL) andstirred at 15° C. for 1 h. Then the mixture was filtered and extractedwith DCM (40 mL×3). The combined organic layers were dried over Na2SO4,filtered and concentrated under reduced pressure. The residue waspurified by column chromatography (SiO2, petroleum ether/ethylacetate=10/1 to 2:1; DCM:MeOH=10:1) to giveN-[(4-methoxyphenyl)methyl]-5-(5-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine(320 mg, 43% yield, 67% purity) as a yellow oil. LC-MS (ESI): m/z (M+H)372.3.

Step D: Synthesis of3-(5-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-amine

A mixture ofN-[(4-methoxyphenyl)methyl]-5-(5-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine(320 mg, 577 μmol, 1 eq) in TFA (7.70 g, 67.5 mmol, 5 mL) was stirred at50° C. for 2 h. After the reaction was completely, the reaction mixturewas concentrated under reduced pressure to give a residue. The residuewas purified by prep-HPLC (column: Luna C18 150×25 5 μm; mobile phase:[water (0.225% FA)-ACN]; B %: 15%-36%, 10 min) to give a crude product.The crude product was diluted with DCM (20 mL)/H2O (20 mL) and adjustedthe pH to 10-12 with NH₃.H2O. Then extracted with DCM (20 mL×3), thecombined organic layers were dried over Na2SO4, filtered andconcentrated under reduced pressure. The residue was purified byprep-TLC (SiO2, DCM:MeOH=10:1) to give5-(5-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine (70 mg, 48% yield)as a white solid. LC-MS (ESI): m/z (M+H) 252.0; 1H NMR (400 MHz,DMSO-d6) 8.46 (s, 1H), 7.88-7.86 (d, 1H), 7.69-7.67 (dd, 1H), 7.64-7.62(d, 2H), 7.57-7.53 (t, 2H), 7.42-7.39 (t, 1H), 6.56 (s, 2H), 2.34 (s,3H).

Step E: Synthesis of2-methyl-N-(3-(5-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-yl)benzamide(Compound R3)

To a solution of 5-(5-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine(70 mg, 279 μmol, 1.0 eq) and pyridine (68.6 mg, 867 μmol, 3.1 eq) inMeCN (9 mL) was added the solution of 2-methyl-benzoyl chloride (60 mg,388 μmol, 50.4 uL, 1.4 eq) in MeCN (1 mL) drop-wise at 80° C. Themixture was stirred at 80° C. for 2 h. Only a few desired product wasformed, then a solution of 2-methylbenzoyl chloride (90 mg, 582.17 μmol,2.1 eq) in MeCN (1 mL) was added dropwise at 80° C. The mixture wasstirred at 80° C. for 16 h. After being cooled to room temperature, thereaction mixture was adjusted pH to 11-12 by saturated aqueous LiOH andstirred at 15° C. for 16 h. The reaction mixture was extracted with DCM(30 mL). The organic layer was washed with saturated aqueous NaHCO3 (30mL), brine (30 mL); dried over Na2SO4, filtered and concentrated underreduced pressure. The residue was purified by prep-HPLC (column:Phenomenex Synergi C18 150×25×10 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 32%-52%, 7.8 min) to give2-methyl-N-[5-(5-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-yl]benzamide(45 mg, 80% yield, HCl salt) as a white solid.

LC-MS (ESI): m/z (M+H) 370.2; 1H NMR (400 MHz, DMSO-d6) 11.32 (s, 1H),8.75-8.74 (m, 1H), 8.22-8.20 (d, 1H), 8.12-8.09 (dd, 1H), 7.84-7.82 (m,2H), 7.64-7.60 (m, 2H), 7.53-7.51 (m, 2H), 7.45-7.40 (td, 1H), 7.33-7.28(m, 2H), 2.46 (s, 3H), 2.20 (s, 3H). 13C NMR (75 MHz, DMSO-d6) 168.6,156.9, 147.8, 146.79, 143.19, 141.8, 136.6, 136.3, 136.0, 134.0, 131.0,130.8, 129.5, 129.2, 127.5, 125.8, 123.8, 122.3, 19.3, 17.9.

Step F: Synthesis ofN-(4-methoxybenzyl)-3-(3-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-amine

A mixture of5-bromo-N-[(4-methoxyphenyl)methyl]-2-phenyl-1,2,4-triazol-3-amine (600mg, 1.62 mmol, 1.0 eq), tributyl-(3-methyl-2-pyridyl)stannane (500 mg,1.31 mmol, 0.8 eq) and [2-(2-aminophenyl)phenyl]-chloro-palladium;dicyclohexyl-[2-(2,6-dimethoxyphenyl)phenyl]phosphane (100 mg, 139 μmol,0.86 eq) in THF (20 mL) was degassed and purged with N2 for three times,and then the mixture was stirred at 90° C. for 60 h under N2. But mostof start material remained, the mixture was filtered and[2-(2-aminophenyl)phenyl]-chloro-palladium;dicyclohexyl-[2-(2,6-dimethoxyphenyl)phenyl]phosphane (100 mg, 139 μmol,0.86 eq) was added. The reaction mixture was stirred at 90° C. foradditional 32 h in a tube. After being cooled to room temperature, thereaction mixture was quenched by addition of saturated aqueous KF (20mL) and stirred at 15° C. for 30 min. Then the mixture was diluted withDCM (20 mL) and extracted with DCM (30 mL×3). The combined organiclayers were dried over Na2SO4, filtered and concentrated under reducedpressure. The residue was purified by column chromatography (TLC: EtOAc;SiO2, petroleum ether/ethyl acetate=5/1 to 2:1; DCM:MeOH=10:1) to give acrude product. The crude product was repurified by prep-TLC (SiO2,EtOAc) to giveN-[(4-methoxyphenyl)methyl]-5-(3-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine(110 mg, 11% yield, 61% purity) as yellow oil. LC-MS (ESI): m/z (M+H)372.0.

Step G: Synthesis of3-(3-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-amine

A mixture ofN-[(4-methoxyphenyl)methyl]-5-(3-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine(110 mg, 181 μmol, 1.0 eq) in TFA (4.62 g, 40.5 mmol, 3 mL) was stirredat 50° C. for 2 h. After completion of the reaction, the reactionmixture was concentrated under reduced pressure. The residue was dilutedwith DCM (15 mL) and water (10 mL), and adjust pH to 9-10 by K2CO3(solid). Then the resulting was extracted with DCM (15 mL×2), thecombined organic layers were concentrated under reduced pressure. Theresidue was purified by prep-TLC (TLC: DCM:MeOH=10:1; SiO2,DCM:MeOH=10:1) to give5-(3-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine (55 mg, crude) asa white solid. LC-MS (ESI): m/z (M+H) 252.0.

Step H: Synthesis of2-methyl-N-(3-(3-methylpyridin-2-yl)-1-phenyl-1H-1,2,4-triazol-5-yl)benzamide(Compound S3)

To a solution of 5-(3-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-amine(55 mg, 219 μmol, 1.0 eq), pyridine (98 mg, 1.24 mmol, 5.7 eq) and DMAP(26 mg, 213 μmol, 0.97 eq) in toluene (9 mL) was added the solution of2-methylbenzoyl chloride (100 mg, 647 mol, 3.0 eq) in toluene (1 mL)dropwise at 80° C. Then the mixture was stirred at 80° C. for 2 h. Thereaction mixture was concentrated under reduced pressure. The residuewas dissolved in CH3CN (10 mL) and adjusted the pH to 11-12 by saturatedaqueous LiOH and stirred at 15° C. for 16 h. The reaction mixture wasextracted with DCM (30 mL). The organic layer was washed with saturatedNaHCO3 (30 mL), brine (30 mL×1), dried over Na2SO4, filtered andconcentrated under reduced pressure. The residue was purified byprep-HPLC (column: Phenomenex Synergi C18 150×25×10 μm; mobile phase:[water (0.05% HCl)-ACN]; B %: 30%-45%, 7.8 min) to give2-methyl-N-[5-(3-methyl-2-pyridyl)-2-phenyl-1,2,4-triazol-3-yl]benzamide(16.1 mg, 17.54% yield, HCl salt) as a white solid. LC-MS (ESI): m/z(M+H) 370.2; 1H NMR (400 MHz, DMSO-d6) 11.30 (s, 1H), 8.69-8.68 (d, 1H),8.26-8.24 (d, 1H), 7.77-7.71 (m, 3H), 7.64-7.61 (m, 2H), 7.57-7.49 (m,2H), 7.45-7.41 (td, 1H), 7.34-7.29 (m, 2H), 2.74 (s, 3H), 2.22 (s, 3H).

Example 14 Synthesis ofN-(1-phenyl-3-(pyridin-2-yl)-1H-1,2,4-triazol-5-yl)benzamide (CompoundY1)

Step A: To a mixture of pyridine-2-carbonitrile (10.0 g, 96.1 mmol) ini-PrOH (57.7 g, 960 mmol, 9.99 eq), NaOMe (155 mg, 2.87 mmol, 0.03 eq)was added at 0° C. Then the mixture was stirred at 20° C. for 4 hr. Themixture was concentrated under vacuum to give a residue. n-Hexane (50mL) and AcOH (0.15 mL) was added to the residue. Then the solution wasfiltered and the filtrate was concentrated under vacuum to giveisopropyl pyridine-2-carboximidate (crude, 2) as brown oil, which wasused for the next step without further purification.

Step B: A solution of NH₂CN (7.5 g, 178 mmol, 1.95 eq), sodiumdihydrogen phosphate (42.0 g, 350 mmol, 3.83 eq) and Na₂HPO₄.12H₂O (32.0g, 89.4 mmol, 0.98 eq) in H₂O (100 mL) was added isopropylpyridine-2-carboximidate (15.0 g crude, 1.0 eq, 2). Then the mixture wasstirred at 20° C. for 16 h. DCM (300 mL) was added to the reactionmixture. The organic layer was concentrated under vacuum. The residuewas purified by flash chromatography (ISCO®; 220 g Sepa Flash® SilicaFlash Column, eluent of 0˜25% Ethyl acetate/Petroleum ether gradient at100 mL/min; TLC (petroleum ether/ethyl acetate=5/1, R_(f)=0.24) to giveisopropyl N-cyanopicolinimidate (10.0 g, 57% yield, 3) as a white solid.LC-MS (ESI): m/z (M+H) 190.1; ¹H NMR (400 MHz, DMSO-d₆) 8.81-8.79 (m,1H), 8.11-8.07 (dt, 1H), 7.95-7.93 (m, 1H), 7.74-7.71 (m, 1H), 5.34-5.28(hept, 1H), 1.41-1.39 (d, 6H).

Step C: A mixture of isopropyl N-cyanopicolinimidate (1.0 g, 5.29 mmol,3) and phenylhydrazine (627 mg, 5.80 mmol, 1.1 eq) in MeOH (15 mL) wasstirred at 80° C. for 16 h. After being cooled to room temperature, thereaction mixture was concentrated under reduced pressure. The residuewas purified by column chromatography (TLC: petroleum ether/ethylacetate=0/1, R_(f)=0.1) (SiO₂, petroleum ether/ethyl acetate=1/0, 5/1,2/1, 0:1) to afford 2-phenyl-5-(2-pyridyl)-1,2,4-triazol-3-amine (900mg, 70% yield, 4) as a brown solid. LC-MS (ESI): m/z (M+H) 238.1.

Step D: To a mixture of 2-phenyl-5-(2-pyridyl)-1,2,4-triazol-3-amine(200 mg, 843 μmol, 1.0 eq, 4) and NaOH (180 mg, 4.50 mmol, 5.34 eq) inTHF (2 mL) and H₂O (2 mL), benzoyl chloride (242 mg, 1.72 mmol, 2.04 eq)was added. Then the mixture was stirred at 20° C. for 20 h. The reactionmixture was adjusted to pH=7 by HCl (2 M in water, 2 mL). The mixturewas concentrated under vacuum. The residue was purified by reversedphase column (HCl condition) to giveN-[2-phenyl-5-(2-pyridyl)-1,2,4-triazol-3-yl]benzamide (70.1 mg, 21%yield, Compound Y1) as a white solid. LC-MS (ESI): m/z (M+H) 342.1; ¹HNMR (400 MHz, DMSO-d₆+D₂O) 8.83-8.82 (d, 1H), 8.44-8.41 (m, 2H),7.97-7.95 (d, 2H), 7.89-7.86 (m, 1H), 7.74-7.72 (m, 2H), 7.64-7.60 (m,1H), 7.55-7.50 (m, 4H), 7.48-7.44 (dd, 1H). ¹³C NMR (100 MHz, DMSO-d₆)167.1, 156.7, 148.8, 146.8, 145.5, 142.9, 137.0, 133.3, 132.5, 130.0,129.6, 129.1, 128.6, 126.6, 123.8, 123.5

Example 15 Synthesis ofN-(3-(pyridin-2-yl)-1-(4-(trifluoromethyl)phenyl)-1H-1,2,4-triazol-5-yl)benzamide(Compound H3)

Step A: To a mixture of (4-(trifluoromethyl)phenyl)hydrazine (260 mg,1.14 mmol, 1.10 eq) in MeOH (10 mL) was added TEA (524 mg, 5.2 mmol,0.72 mL, 5.00 eq) and isopropyl-N-cyanopicolinimidate (200 mg, 1.04mmol, 1.0 eq, 3 from Example 14). The resultant mixture was stirred at80° C. for 4 h. After being cooled to room temperature, the reactionmixture was concentrated under vacuum. The residue was purified byprep-TLC (DCM/MeOH=10/1, R_(f)=0.24).3-(pyridin-2-yl)-1-(4-(trifluoromethoxy)phenyl)-1H-1,2,4-triazol-5-amine(418 mg, crude, 5) was obtained as yellow oil. LC-MS (ESI): m/z (M+H)321.9; ¹H NMR (400 MHz, DMSO-d₆) 8.62 (s, 1H), 7.97-7.96 (m, 1H), 7.87(s, 1H), 7.77-7.75 (m, 2H), 7.56-7.54 (m, 2H), 7.40 (s, 1H), 6.70 (s,2H).

Step B: To a solution of5-(2-pyridyl)-2-[4-(trifluoromethoxy)phenyl]-1,2,4-triazol-3-amine (150mg, 467 μmol, 1 eq) in CH₃CN (15 mL) was added pyridine (185 mg, 2.33mmol, 5.0 eq) and 2-methylbenzoyl chloride (86 mg, 556 μmol, 1.19 eq).The resulting solution was allowed to stir at 75° C. for 48 h. Afterbeing cooled to room temperature, the reaction mixture was concentratedunder reduced pressure. The residue was purified by prep-HPLC (HClcondition). The desired fractions were collected and the solvent wasremoved by lyophilization to afford2-methyl-N-[5-(2-pyridyl)-2-[4-(trifluoromethoxy)phenyl]-1,2,4-triazol-3-yl]benzamide(102 mg, 45% yield, 98.4% purity, HCl salt, Compound H3 HCl) as a whitesolid. LC-MS (ESI): m/z (M+H) 440.2; ¹H NMR (400 MHz, DMSO-d₆) 11.32 (s,1H), 8.75-8.73 (m, 1H), 7.83-7.81 (m, 1H), 8.12-8.08 (m, 1H), 7.84-7.80(m, 2H), 7.63-7.60 (m, 3H), 7.52-7.50 (m, 1H), 7.44-7.40 (m, 1H),7.33-7.28 (m, 2H), 2.17 (s, 3H). ¹³C NMR (100 MHz, DMSO-d₆) 168.5,157.9, 148.2, 148.0, 147.9, 146.6, 140.2, 136.3, 135.7, 133.9, 131.0,130.8, 127.5, 125.9, 125.8, 125.5, 122.5, 122.2, 118.7, 19.1.

Example 16 Synthesis of2-methyl-N-(1-phenyl-3-(pyridin-2-yl)-1H-1,2,4-triazol-5-yl)benzamide(Compound Q1)

Step A: A mixture of 2-methylbenzoic acid (120 mg, 881 μmol, 6) in SOCl₂(10 mL) was stirred at 80° C. for 2 h. The mixture was concentratedunder vacuum to give 2-methylbenzoyl chloride (130 mg, crude, 7) ascolorless oil, which was used for the next step without furtherpurification.

Step B: To a mixture of 2-phenyl-5-(2-pyridyl)-1,2,4-triazol-3-amine(200 mg, 826 μmol, 4 from Example 14) and aqueous NaOH (2 M, 4.84 eq) inTHF (2 mL), 2-methylbenzoyl chloride (130 mg crude, 1.02 eq, 7) wasadded. Then the mixture was stirred at 20° C. for 20 h. The reactionmixture was adjusted to pH=7 by conc. hydrochloric acid (12 M, 0.2 mL).The mixture was extracted with DCM (50 mL) and washed with water (15mL). The organic layer was concentrated under reduced pressure. Theresidue was purified by prep-HPLC (HCl condition) to afford2-methyl-N-[2-phenyl-5-(2-pyridyl)-1,2,4-triazol-3-yl]benzamide (HClsalt, 35.1 mg, 11% yield, Compound Q1 HCl) as a pink solid. LC-MS (ESI):m/z (M+H) 356.2; ¹H NMR (400 MHz, DMSO-d6) 11.41 (s, 1H), 8.81-8.79 (d,1H), 8.34-8.29 (m, 2H), 7.70-7.77 (m, 1H), 7.72-7.70 (m, 2H), 7.62-7.58(dd, 2H), 7.55-7.50 (m, 2H), 7.43-7.39 (dd, 1H), 7.32-7.26 (m, 2H), 2.20(s, 3H). ¹³C NMR (100 MHz, DMSO-d₆) 169.0, 157.1, 148.4, 147.2, 145.9,142.5, 137.0, 134.3, 131.4, 131.3, 130.0, 129.8, 128.0, 126.4, 126.3,124.3, 123.3, 19.7.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.

While the invention has been described in connection with proposedspecific embodiments thereof, it will be understood that it is capableof further modifications and this application is intended to cover anyvariations, uses, or adaptations of the invention following, in general,the principles of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth and as follows in the scope ofthe appended claims.

What is claimed is:
 1. A compound of formula (IA):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴ —; R¹ and R² are each phenyl, substituted with one or moreR^(5a); R³ is phenyl, optionally substituted with one or more R^(5b); R⁴is each independently H or C1-C3 alkyl; R^(5a) is each independently I,Br, Cl, F, C1-C6 alkyl, C1-C3 haloalkyl, —(C1-C6)-O—(C1-C6), C1-C3alkoxy, C1-C3 haloalkoxy, OH, or COOH; R^(5b) is each independently I,Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH,OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ iseach independently alkyl or haloalkyl.
 2. The compound of claim 1wherein the compound has the structure of formula (IB):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each phenyl, substituted with one or moreR^(5a); R³ is phenyl, optionally substituted with one or more R^(5b); R⁴is each independently H or C1-C3 alkyl; R^(5a) is each independentlyC1-C6 alkyl; R^(5b) is each independently I, Br, Cl, F, CN, CONH₂,CONHR⁶, CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶,oxo, R⁶, SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ is each independentlyalkyl or haloalkyl.
 3. The compound of claim 1, wherein the compound hasthe structure of formula (IC):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each phenyl, substituted with one or moreR^(5a), wherein at least one of R¹ and R²is

R³ is phenyl, optionally substituted with one or more R^(5b); R⁴ is eachindependently H or C1-C3 alkyl; R^(5a) is each independently I, Br, Cl,F, C1-C6 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, OH, orCOOH; R^(5b) is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶,CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶,SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ is each independently alkyl orhaloalkyl.
 4. The compound of claim 1, wherein the compound has thestructure of formula (ID):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); R⁴ is eachindependently H or C1-C3 alkyl; R^(5b) is each independently I, Br, Cl,F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶,—COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ is eachindependently alkyl or haloalkyl.
 5. The compound of claim 1, whereinthe compound has the structure of formula (IE):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L ³is a bond; M¹ and M² are each —NHC(O)—; R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); and R^(5b)is each independently I, Br, Cl, F, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3alkoxy, C1-C3 haloalkoxy, OH, or COOH.
 6. The compound of claim 1,wherein the compound has the structure of formula (IG):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each —NHC(O)—; R¹ and R² are each

R³ is phenyl; and R^(5a) a is each independently C1-C6 alkyl, C1-C3haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, OH, or COOH.
 7. The compoundof claim 5, wherein the compound has the structure of formula (IF):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each —NHC(O)—; R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); and R^(5b)is each independently C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3haloalkoxy, OH, or COOH.
 8. A compound having one of the followingstructures:

or a pharmaceutically acceptable salt or solvate thereof.
 9. Thecompound of claim 8 having the following structure:

or a pharmaceutically acceptable salt or solvate thereof.
 10. Apharmaceutical composition comprising a pharmaceutically acceptablecarrier or a pharmaceutically acceptable excipient and a compound ofclaim 1, wherein the compound has the structure of formula (IA):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each phenyl, substituted with one or moreR^(5a); R³ is phenyl, optionally substituted with one or more R^(5b); R⁴is each independently H or C1-C3 alkyl; R^(5a) is each independently I,Br, Cl, F, C1-C6 alkyl, C1-C3 haloalkyl, —(C1-C6)-O—(C1-C6), C1-C3alkoxy, C1-C3 haloalkoxy, OH, or COOH; R^(5b) is each independently I,Br, Cl, F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH,OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ iseach independently alkyl or haloalkyl.
 11. The pharmaceuticalcomposition of claim 10, wherein the compound has the structure offormula (IB):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each phenyl, substituted with one or moreR^(5a); R³ is phenyl, optionally substituted with one or more R^(5b); R⁴is each independently H or C1-C3 alkyl; R^(5a) is each independentlyC1-C6 alkyl; R^(5b) is each independently I, Br, Cl, F, CN, CONH₂,CONHR⁶, CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶,oxo, R⁶, SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ is each independentlyalkyl or haloalkyl.
 12. The pharmaceutical composition of claim 10,wherein the compound has the structure of formula (IC):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each phenyl, substituted with one or moreR^(5a), wherein at least one of R¹ and R² is

R³ is phenyl, optionally substituted with one or more R^(5b); R⁴ is eachindependently H or C1-C3 alkyl; R^(5a) is each independently I, Br, Cl,F, C1-C6 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, OH, orCOOH; R^(5b)is each independently I, Br, Cl, F, CN, CONH₂, CONHR⁶,CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OH, OR⁶, —COOR⁶, OSO₃R⁶, oxo, R⁶,SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ is each independently alkyl orhaloalkyl.
 13. The pharmaceutical composition of claim 10, wherein thecompound has the structure of formula (ID):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³ isa bond; M¹ and M² are each independently selected from —NR⁴C(O)— or—C(O)NR⁴—; R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); R⁴ is eachindependently H or C1-C3 alkyl; R^(5b) is each independently I, Br, Cl,F, CN, CONH₂, CONHR⁶, CONR⁶R⁶, COOH, NH₂, NHR⁶, NO₂, NR⁶R⁶, OR⁶, —COOR⁶,OSO₃R⁶, oxo, R⁶, SH, SO₂R⁶, SO₃H, SO₃R⁶, or SR⁶; and R⁶ is eachindependently alkyl or haloalkyl.
 14. The pharmaceutical composition ofclaim 10, wherein the compound has the structure of formula (IE):

or a pharmaceutically acceptable salt or solvate thereof, wherein: L³isa bond; M¹ and M² are each —NHC(O)—; R¹ and R² are each

R³ is phenyl, optionally substituted with one or more R^(5b); and R^(5b)is each independently I, Br, Cl, F, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3alkoxy, C1-C3 haloalkoxy, OH, or COOH.
 15. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier or a pharmaceuticallyacceptable excipient and a compound of any compound claim
 8. 16. Thepharmaceutical composition of claim 10, wherein the compound has thefollowing structure:

or a pharmaceutically acceptable salt or solvate thereof.